Full-length expressed genetic markers

ABSTRACT

The invention provides full-length expressed genetic markers (FLEXGEM) and polynucleotides which identify and encode FLEXGEM. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with expression of FLEXGEM.

[0001] This application is a continuation application of U.S. application Ser. No. 09/311,894, filed May 14, 1999.

FIELD OF THE INVENTION

[0002] The present invention relates to nucleic acid and amino acid sequences of full-length expressed genetic markers and to the use of these sequences in the analysis of human gene expression, genetic linkage, and genetic variability, and in the diagnosis and treatment of diseases linked to altered gene expression.

BACKGROUND OF THE INVENTION

[0003] It is estimated that only 2% of mammalian DNA encodes proteins, and only a small fraction of the genes that encode proteins is actually expressed in a particular cell at any time. The various types of cells in a multicellular organism differ dramatically both in structure and function, and the identity of a particular cell is conferred by its unique pattern of gene expression. In addition, different cell types express overlapping but distinctive sets of genes throughout development. Cell growth and proliferation, cell differentiation, the immune response, apoptosis, and other processes that contribute to organism development and survival are governed by regulation of gene expression. Appropriate gene regulation also ensures that cells function efficiently by expressing only those genes whose functions are required at a given time. Factors that influence gene expression include extracellular signals that mediate cell-cell communication and coordinate the activities of different cell types. Gene expression is regulated at the level of DNA and RNA transcription, and at the level of mRNA translation.

[0004] Aberrant expression or mutations in genes and their products may cause, or increase susceptibility to, a variety of human diseases such as cancer and other cell proliferative disorders. The identification of these genes and their products is the basis of an ever-expanding effort to finding markers for early detection of diseases and targets for their prevention and treatment. For example, cancer represents a type of cell proliferative disorder that affects nearly every tissue in the body. The development of cancer, or oncogenesis, is often correlated with the conversion of a normal gene into a cancer-causing gene, or oncogene, through abnormal expression or mutation. Oncoproteins, the products of oncogenes, include a variety of molecules that influence cell proliferation, such as growth factors, growth factor receptors, intracellular signal transducers, nuclear transcription factors, and cell-cycle control proteins. In contrast, tumor-suppressor genes are involved in inhibiting cell proliferation. Mutations which reduce or abrogate the function of tumor-suppressor genes result in aberrant cell proliferation and cancer. Thus a wide variety of genes and their products have been found that are associated with cell proliferative disorders such as cancer, but many more may exist that are yet to be discovered.

[0005] DNA-based arrays can provide an efficient, high-throughput method to examine gene expression and genetic variability. For example, SNPs, or single nucleotide polymorphisms, are the most common type of human genetic variation. DNA-based arrays can dramatically accelerate the discovery of SNPs in hundreds and even thousands of genes. Likewise, such arrays can be used for SNP genotyping in which DNA samples from individuals or populations are assayed for the presence of selected SNPs. These approaches will ultimately lead to the systematic identification of all genetic variations in the human genome and the correlation of certain genetic variations with disease susceptibility, responsiveness to drug treatments, and other medically relevant information. (See, for example, Wang, D. G. et al. (1998) Science 280:1077-1082.)

[0006] DNA-based array technology is especially important for the rapid analysis of global gene expression patterns. For example, genetic predisposition, disease, or therapeutic treatment may directly or indirectly affect the expression of a large number of genes in a given tissue. In this case, it is useful to develop a profile, or transcript image, of all the genes that are expressed and the levels at which they are expressed in that particular tissue. A profile generated from an individual or population affected with a certain disease or undergoing a particular therapy may be compared with a profile likewise generated from a control individual or population. Such analysis does not require knowledge of gene function, as the expression profiles can subjected to mathematical analyses which simply treat each gene as a marker. Furthermore, gene expression profiles may help dissect biological pathways by identifying all the genes expressed, for example, at a certain developmental stage, in a particular tissue, or in response to disease or treatment. (See, for example, Lander, E. S. et al. (1996) Science 274:536-539.)

[0007] The discovery of new human full-length expressed genetic markers and the polynucleotides encoding them satisfies a need in the art by providing new compositions which are useful in the diagnosis, prevention, and treatment of developmental, cell proliferative, and immunological disorders.

SUMMARY OF THE INVENTION

[0008] The invention features substantially purified polypeptides, human full-length expressed genetic markers, referred to collectively as “FLEXGEM” and individually as “FLEXGEM-1,” “FLEXGEM-2,” “FLEXGEM-3,” “FLEXGEM-4,” “FLEXGEM-5,” “FLEXGEM-6,” “FLEXGEM-7,” “FLEXGEM-8,” “FLEXGEM-9,” “FLEXGEM-10,” “FLEXGEM-11,” “FLEXGEM-12,” “FLEXGEM-13,” “FLEXGEM-14,” “FLEXGEM-15,” “FLEXGEM-16,” “FLEXGEM-17, ” “FLEXGEM-18,” “FLEXGEM-19,” and “FLEXGEM-20.” In one aspect, the invention provides a substantially purified polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 (SEQ ID NO:1-20), and fragments thereof.

[0009] The invention also provides an isolated and purified polynucleotide encoding the polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-20 and fragments thereof. The invention further provides an isolated and purified polynucleotide having a sequence which is complementary to the polynucleotide encoding the polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-20 and fragments thereof.

[0010] Additionally, the invention provides a method for detecting a polynucleotide in a sample containing nucleic acids, the method comprising the steps of (a) hybridizing the complement of the polynucleotide sequence to at least one of the polynucleotides of the sample, thereby forming a hybridization complex; and (b) detecting the hybridization complex, wherein the presence of the hybridization complex correlates with the presence of a polynucleotide in the sample. In one aspect, the method further comprises amplifying the polynucleotide prior to hybridization.

[0011] The invention further provides an expression vector containing at least a fragment of the polynucleotide encoding the polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-20 and fragments thereof. In another aspect, the expression vector is contained within a host cell.

[0012] The invention also provides a method for producing a polypeptide, the method comprising the steps of: (a) culturing the host cell containing an expression vector containing at least a fragment of a polynucleotide under conditions suitable for the expression of the polypeptide; and (b) recovering the polypeptide from the host cell culture.

[0013] The invention also provides an isolated and purified polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40 (SEQ ID NO:21-40), and fragments thereof. The invention further provides an isolated and purified polynucleotide having a sequence which is complementary to the polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:21-40 and fragments thereof.

[0014] The invention also provides a composition for the detection of altered expression of human FLEXGEM polynucleotides comprising at least one of these complementary polynucleotides and a detectable label. Also provided is an isolated and purified polynucleotide which is complementary to at least a fragment of a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:21-40 and fragments thereof.

[0015] The invention further encompasses a microarray containing at least a fragment of at least one of the polynucleotides comprising the polynucleotide sequence selected from the group consisting of SEQ ID NO:21-40, or fragments thereof. Also encompassed is a method for generating a transcript image of a sample containing nucleic acids, the method comprising the steps of hybridizing the polynucleotides on the microarray containing at least one of the polynucleotides selected from the group comprising SEQ ID NO:21-40, or fragments thereof, to labeled complementary polynucleotides from the sample under conditions suitable for the formation of a hybridization complex, and quantifying the expression level of the polynucleotides encoding the polypeptides in the biological sample.

[0016] The invention also provides a pharmaceutical composition comprising a substantially purified polypeptide having the amino acid sequence selected from the group consisting of SEQ ID NO:1-20 and fragments thereof, in conjunction with a suitable pharmaceutical carrier.

[0017] The invention further includes a purified antibody which binds to a polypeptide selected from the group consisting of SEQ ID NO:1-20 and fragments thereof. The invention also provides a purified agonist and a purified antagonist to the polypeptide.

[0018] The invention also provides a method for treating or preventing a disorder associated with decreased expression or activity of FLEXGEM, the method comprising administering to a subject in need of such treatment an effective amount of a pharmaceutical composition comprising a substantially purified polypeptide having the amino acid sequence selected from the group consisting of SEQ ID NO:1-20 and fragments thereof, in conjunction with a suitable pharmaceutical carrier.

[0019] The invention also provides a method for treating or preventing a disorder associated with increased expression or activity of FLEXGEM, the method comprising administering to a subject in need of such treatment an effective amount of an antagonist of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-20 and fragments thereof.

[0020] The invention further provides a method for identifying a test compound which specifically binds to a polypeptide selected from the group consisting of SEQ ID NO:1-20, the method comprising the steps of providing a test compound, combining the polypeptide with the test compound under suitable conditions for a time sufficient to allow binding, and detecting the binding of the polypeptide to the test compound.

BRIEF DESCRIPTION OF THE TABLES

[0021] Table 1 shows polypeptide and nucleotide sequence identification numbers (SEQ ID NOs), clone identification numbers (clone IDs), cDNA libraries, and cDNA fragments used to assemble full-length sequences encoding FLEXGEM.

[0022] Table 2 shows features of each polypeptide sequence, including potential phosphorylation and glycosylation sites.

[0023] Table 3 shows useful fragments of each nucleic acid sequence; the tissue-specific expression patterns of each nucleic acid sequence as determined by northern analysis; diseases, disorders, or conditions associated with these tissues; and the vector into which each cDNA was cloned.

[0024] Table 4 describes the tissues used to construct the cDNA libraries from which cDNA clones encoding FLEXGEM were isolated.

[0025] Table 5 shows the tools, programs, and algorithms used to analyze FLEXGEM, along with applicable descriptions, references, and threshold parameters.

DESCRIPTION OF THE INVENTION

[0026] Before the present proteins, nucleotide sequences, and methods are described, it is understood that this invention is not limited to the particular machines, materials and methods described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

[0027] It must be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to “a host cell” includes a plurality of such host cells, and a reference to “an antibody” is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.

[0028] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any machines, materials, and methods similar or equivalent to those described herein can be used to practice or test the present invention, the preferred machines, materials and methods are now described. All publications mentioned herein are cited for the purpose of describing and disclosing the cell lines, protocols, reagents and vectors which are reported in the publications and which might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

Definitions

[0029] “FLEXGEM” refers to the amino acid sequences of substantially purified FLEXGEM obtained from any species, particularly a mammalian species, including bovine, ovine, porcine, murine, equine, and preferably the human species, from any source, whether natural, synthetic, semi-synthetic, or recombinant.

[0030] The term “agonist” refers to a molecule which, when bound to FLEXGEM, increases or prolongs the duration of the effect of FLEXGEM. Agonists may include proteins, nucleic acids, carbohydrates, or any other molecules which bind to and modulate the effect of FLEXGEM.

[0031] An “allelic variant” is an alternative form of the gene encoding FLEXGEM. Allelic variants may result from at least one mutation in the nucleic acid sequence and may result in altered mRNAs or in polypeptides whose structure or function may or may not be altered. Any given natural or recombinant gene may have none, one, or many allelic forms. Common mutational changes which give rise to allelic variants are generally ascribed to natural deletions, additions, or substitutions of nucleotides. Each of these types of changes may occur alone, or in combination with the others, one or more times in a given sequence.

[0032] “Altered” nucleic acid sequences encoding FLEXGEM include those sequences with deletions, insertions, or substitutions of different nucleotides, resulting in a polynucleotide the same as FLEXGEM or a polypeptide with at least one functional characteristic of FLEXGEM. Included within this definition are polymorphisms which may or may not be readily detectable using a particular oligonucleotide probe of the polynucleotide encoding FLEXGEM, and improper or unexpected hybridization to allelic variants, with a locus other than the normal chromosomal locus for the polynucleotide sequence encoding FLEXGEM. The encoded protein may also be “altered,” and may contain deletions, insertions, or substitutions of amino acid residues which produce a silent change and result in a functionally equivalent FLEXGEM. Deliberate amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues, as long as the biological or immunological activity of FLEXGEM is retained. For example, negatively charged amino acids may include aspartic acid and glutamic acid, positively charged amino acids may include lysine and arginine, and amino acids with uncharged polar head groups having similar hydrophilicity values may include leucine, isoleucine, and valine; glycine and alanine; asparagine and glutamine; serine and threonine; and phenylalanine and tyrosine.

[0033] The terms “amino acid” and “amino acid sequence” refer to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and to naturally occurring or synthetic molecules. In this context, “fragments,” “immunogenic fragments,” or “antigenic fragments” refer to fragments of FLEXGEM which are preferably at least 5 to about 15 amino acids in length, most preferably at least 14 amino acids, and which retain some biological activity or immunological activity of FLEXGEM. Where “amino acid sequence” is recited to refer to an amino acid sequence of a naturally occurring protein molecule, “amino acid sequence” and like terms are not meant to limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule.

[0034] “Amplification” relates to the production of additional copies of a nucleic acid sequence. Amplification is generally carried out using polymerase chain reaction (PCR) technologies well known in the art.

[0035] The term “antagonist” refers to a molecule which, when bound to FLEXGEM, decreases the amount or the duration of the effect of the biological or immunological activity of FLEXGEM. Antagonists may include proteins, nucleic acids, carbohydrates, antibodies, or any other molecules which decrease the effect of FLEXGEM.

[0036] The term “antibody” refers to intact molecules as well as to fragments thereof, such as Fab, F(ab′)₂, and Fv fragments, which are capable of binding the epitopic determinant. Antibodies that bind FLEXGEM polypeptides can be prepared using intact polypeptides or using fragments containing small peptides of interest as the immunizing antigen. The polypeptide or oligopeptide used to immunize an animal (e.g., a mouse, a rat, or a rabbit) can be derived from the translation of RNA, or synthesized chemically, and can be conjugated to a carrier protein if desired. Commonly used carriers that are chemically coupled to peptides include bovine serum albumin, thyroglobulin, and keyhole limpet hemocyanin (KLH). The coupled peptide is then used to immunize the animal.

[0037] The term “antigenic determinant” refers to that fragment of a molecule (i.e., an epitope) that makes contact with a particular antibody. When a protein or a fragment of a protein is used to immunize a host animal, numerous regions of the protein may induce the production of antibodies which bind specifically to antigenic determinants (given regions or three-dimensional structures on the protein). An antigenic determinant may compete with the intact antigen (i.e., the immunogen used to elicit the immune response) for binding to an antibody.

[0038] The term “antisense” refers to any composition containing a nucleic acid sequence which is complementary to the “sense” strand of a specific nucleic acid sequence. Antisense molecules may be produced by any method including synthesis or transcription. Once introduced into a cell, the complementary nucleotides combine with natural sequences produced by the cell to form duplexes and to block either transcription or translation. The designation “negative” can refer to the antisense strand, and the designation “positive” can refer to the sense strand.

[0039] The term “biologically active” refers to a protein having structural, regulatory, or biochemical functions of a naturally occurring molecule. Likewise, “immunologically active” refers to the capability of the natural, recombinant, or synthetic FLEXGEM, or of any oligopeptide thereof, to induce a specific immune response in appropriate animals or cells and to bind with specific antibodies.

[0040] The terms “complementary” and “complementarity” refer to the natural binding of polynucleotides by base pairing. For example, the sequence “5′ A-G-T 3′” bonds to the complementary sequence “3′ T-C-A 5′.” Complementarity between two single-stranded molecules may be “partial,” such that only some of the nucleic acids bind, or it may be “complete,” such that total complementarity exists between the single stranded molecules. The degree of complementarity between nucleic acid strands has significant effects on the efficiency and strength of the hybridization between the nucleic acid strands. This is of particular importance in amplification reactions, which depend upon binding between nucleic acids strands, and in the design and use of peptide nucleic acid (PNA) molecules.

[0041] A “composition comprising a given polynucleotide sequence” and a “composition comprising a given amino acid sequence” refer broadly to any composition containing the given polynucleotide or amino acid sequence. The composition may comprise a dry formulation or an aqueous solution. Compositions comprising polynucleotide sequences encoding FLEXGEM or fragments of FLEXGEM may be employed as hybridization probes. The probes may be stored in freeze-dried form and may be associated with a stabilizing agent such as a carbohydrate. In hybridizations, the probe may be deployed in an aqueous solution containing salts (e.g., NaCl), detergents (e.g., sodium dodecyl sulfate; SDS), and other components (e.g., Denhardt's solution, dry milk, salmon sperm DNA, etc.).

[0042] “Consensus sequence” refers to a nucleic acid sequence which has been resequenced to resolve uncalled bases, extended using the XL-PCR kit (Perkin-Elmer, Norwalk Conn.) in the 5′ and/or the 3′ direction, and resequenced, or which has been assembled from the overlapping sequences of more than one Incyte Clone using a computer program for fragment assembly, such as the GELVIEW fragment assembly system (GCG, Madison Wis.). Some sequences have been both extended and assembled to produce the consensus sequence.

[0043] The term “correlates with expression of a polynucleotide” indicates that the detection of the presence of nucleic acids, the same or related to a nucleic acid sequence encoding FLEXGEM, by northern analysis is indicative of the presence of nucleic acids encoding FLEXGEM in a sample, and thereby correlates with expression of the transcript from the polynucleotide encoding FLEXGEM.

[0044] A “deletion” refers to a change in the amino acid or nucleotide sequence that results in the absence of one or more amino acid residues or nucleotides.

[0045] The term “derivative” refers to the chemical modification of a polypeptide sequence, or a polynucleotide sequence. Chemical modifications of a polynucleotide sequence can include, for example, replacement of hydrogen by an alkyl, acyl, or amino group. A derivative polynucleotide encodes a polypeptide which retains at least one biological or immunological function of the natural molecule. A derivative polypeptide is one modified by glycosylation, pegylation, or any similar process that retains at least one biological or immunological function of the polypeptide from which it was derived.

[0046] A “full length” nucleotide sequence is one containing at least a start site for translation to a protein sequence, followed by an open reading frame and a stop site, and encoding a “full length” polypeptide.

[0047] The term “similarity” refers to a degree of complementarity. There may be partial similarity or complete similarity. The word “identity” may substitute for the word “similarity.” A partially complementary sequence that at least partially inhibits an identical sequence from hybridizing to a target nucleic acid is referred to as “substantially similar.” The inhibition of hybridization of the completely complementary sequence to the target sequence may be examined using a hybridization assay (Southern or northern blot, solution hybridization, and the like) under conditions of reduced stringency. A substantially similar sequence or hybridization probe will compete for and inhibit the binding of a completely similar (identical) sequence to the target sequence under conditions of reduced stringency. This is not to say that conditions of reduced stringency are such that non-specific binding is permitted, as reduced stringency conditions require that the binding of two sequences to one another be a specific (i.e., a selective) interaction. The absence of non-specific binding may be tested by the use of a second target sequence which lacks even a partial degree of complementarity (e.g., less than about 30% similarity or identity). In the absence of non-specific binding, the substantially similar sequence or probe will not hybridize to the second non-complementary target sequence.

[0048] The phrases “percent identity” and “% identity” refer to the percentage of sequence similarity found in a comparison of two or more amino acid or nucleic acid sequences. Percent identity can be determined electronically, e.g., by using the MEGALIGN program (DNASTAR, Madison Wis.) which creates alignments between two or more sequences according to methods selected by the user, e.g., the clustal method. (See, e.g., Higgins, D. G. and P. M. Sharp (1988) Gene 73:237-244.) Parameters for each method may be the default parameters provided by MEGALIGN or may be specified by the user. The clustal algorithm groups sequences into clusters by examining the distances between all pairs. The clusters are aligned pairwise and then in groups. The percentage similarity between two amino acid sequences, e.g., sequence A and sequence B, is calculated by dividing the length of sequence A, minus the number of gap residues in sequence A, minus the number of gap residues in sequence B, into the sum of the residue matches between sequence A and sequence B, times one hundred. Gaps of low or of no similarity between the two amino acid sequences are not included in determining percentage similarity. Percent identity between nucleic acid sequences can also be counted or calculated by other methods known in the art, e.g., the Jotun Hein method. (See, e.g., Hein, J. (1990) Methods Enzymol. 183:626-645.) Identity between sequences can also be determined by other methods known in the art, e.g., by varying hybridization conditions.

[0049] “Human artificial chromosomes” (HACs) are linear microchromosomes which may contain DNA sequences of about 6 kb to 10 Mb in size, and which contain all of the elements required for stable mitotic chromosome segregation and maintenance.

[0050] The term “humanized antibody” refers to antibody molecules in which the amino acid sequence in the non-antigen binding regions has been altered so that the antibody more closely resembles a human antibody, and still retains its original binding ability.

[0051] “Hybridization” refers to any process by which a strand of nucleic acid binds with a complementary strand through base pairing.

[0052] The term “hybridization complex” refers to a complex formed between two nucleic acid sequences by virtue of the formation of hydrogen bonds between complementary bases. A hybridization complex may be formed in solution (e.g., C₀t or R₀t analysis) or formed between one nucleic acid sequence present in solution and another nucleic acid sequence immobilized on a solid support (e.g., paper, membranes, filters, chips, pins or glass slides, or any other appropriate substrate to which cells or their nucleic acids have been fixed).

[0053] The words “insertion” and “addition” refer to changes in an amino acid or nucleotide sequence resulting in the addition of one or more amino acid residues or nucleotides, respectively, to the sequence found in the naturally occurring molecule.

[0054] “Immune response” can refer to conditions associated with inflammation, trauma, immune disorders, or infectious or genetic disease, etc. These conditions can be characterized by expression of various factors, e.g., cytokines, chemokines, and other signaling molecules, which may affect cellular and systemic defense systems.

[0055] The term “microarray” refers to an arrangement of distinct polynucleotides on a substrate.

[0056] The terms “element” and “array element” in a microarray context, refer to hybridizable polynucleotides arranged on the surface of a substrate.

[0057] The term “modulate” refers to a change in the activity of FLEXGEM. For example, modulation may cause an increase or a decrease in protein activity, binding characteristics, or any other biological, functional, or immunological properties of FLEXGEM.

[0058] The phrases “nucleic acid” and “nucleic acid sequence” refer to a nucleotide, oligonucleotide, polynucleotide, or any fragment thereof. These phrases also refer to DNA or RNA of genomic or synthetic origin which may be single-stranded or double-stranded and may represent the sense or the antisense strand, to peptide nucleic acid (PNA), or to any DNA-like or RNA-like material. In this context, “fragments” refers to those nucleic acid sequences which, when translated, would produce polypeptides retaining some functional characteristic, e.g., antigenicity, or structural domain characteristic, e.g., ATP-binding site, of the full-length polypeptide.

[0059] The terms “operably associated” and “operably linked” refer to functionally related nucleic acid sequences. A promoter is operably associated or operably linked with a coding sequence if the promoter controls the translation of the encoded polypeptide. While operably associated or operably linked nucleic acid sequences can be contiguous and in the same reading frame, certain genetic elements, e.g., repressor genes, are not contiguously linked to the sequence encoding the polypeptide but still bind to operator sequences that control expression of the polypeptide.

[0060] The term “oligonucleotide” refers to a nucleic acid sequence of at least about 6 nucleotides to 60 nucleotides, preferably about 15 to 30 nucleotides, and most preferably about 20 to 25 nucleotides, which can be used in PCR amplification or in a hybridization assay or microarray. “Oligonucleotide” is substantially equivalent to the terms “amplimer,” “primer,” “oligomer,” and “probe,” as these terms are commonly defined in the art.

[0061] “Peptide nucleic acid” (PNA) refers to an antisense molecule or anti-gene agent which comprises an oligonucleotide of at least about 5 nucleotides in length linked to a peptide backbone of amino acid residues ending in lysine. The terminal lysine confers solubility to the composition. PNAs preferentially bind complementary single stranded DNA or RNA and stop transcript elongation, and may be pegylated to extend their lifespan in the cell.

[0062] The term “sample” is used in its broadest sense. A sample suspected of containing nucleic acids encoding FLEXGEM, or fragments thereof, or FLEXGEM itself, may comprise a bodily fluid; an extract from a cell, chromosome, organelle, or membrane isolated from a cell; a cell; genomic DNA, RNA, or cDNA, in solution or bound to a substrate; a tissue; a tissue print; etc.

[0063] The terms “specific binding” and “specifically binding” refer to that interaction between a protein or peptide and an agonist, an antibody, or an antagonist. The interaction is dependent upon the presence of a particular structure of the protein, e.g., the antigenic determinant or epitope, recognized by the binding molecule. For example, if an antibody is specific for epitope “A,” the presence of a polypeptide containing the epitope A, or the presence of free unlabeled A, in a reaction containing free labeled A and the antibody will reduce the amount of labeled A that binds to the antibody.

[0064] The term “stringent conditions” refers to conditions which permit hybridization between polynucleotides and the claimed polynucleotides. Stringent conditions can be defined by salt concentration, the concentration of organic solvent, e.g., formamide, temperature, and other conditions well known in the art. In particular, stringency can be increased by reducing the concentration of salt, increasing the concentration of formamide, or raising the hybridization temperature.

[0065] The term “substantially purified” refers to nucleic acid or amino acid sequences that are removed from their natural environment and are isolated or separated, and are at least about 60% free, preferably about 75% free, and most preferably about 90% free from other components with which they are naturally associated.

[0066] A “substitution” refers to the replacement of one or more amino acids or nucleotides by different amino acids or nucleotides, respectively.

[0067] “Substrate” refers to any suitable rigid or semi-rigid support including membranes, filters, chips, slides, wafers, fibers, magnetic or nonmagnetic beads, gels, tubing, plates, polymers, microparticles and capillaries. The substrate can have a variety of surface forms, such as wells, trenches, pins, channels and pores, to which polynucleotides or polypeptides are bound.

[0068] A “transcript image” refers to the collective pattern of gene expression by a particular tissue or cell type under given conditions at a given time.

[0069] “Transformation” describes a process by which exogenous DNA enters and changes a recipient cell. Transformation may occur under natural or artificial conditions according to various methods well known in the art, and may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method for transformation is selected based on the type of host cell being transformed and may include, but is not limited to, viral infection, electroporation, heat shock, lipofection, and particle bombardment. The term “transformed” cells includes stably transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome, as well as transiently transformed cells which express the inserted DNA or RNA for limited periods of time.

[0070] A “variant” of FLEXGEM polypeptides refers to an amino acid sequence that is altered by one or more amino acid residues. The variant may have “conservative” changes, wherein a substituted amino acid has similar structural or chemical properties (e.g., replacement of leucine with isoleucine). More rarely, a variant may have “nonconservative” changes (e.g., replacement of glycine with tryptophan). Analogous minor variations may also include amino acid deletions or insertions, or both. Guidance in determining which amino acid residues may be substituted, inserted, or deleted without abolishing biological or immunological activity may be found using computer programs well known in the art, for example, LASERGENE software (DNASTAR).

[0071] The term “variant,” when used in the context of a polynucleotide sequence, may encompass a polynucleotide sequence related to FLEXGEM. This definition may also include, for example, “allelic” (as defined above), “splice,” “species,” or “polymorphic” variants. A splice variant may have significant identity to a reference molecule, but will generally have a greater or lesser number of polynucleotides due to alternate splicing of exons during mRNA processing. The corresponding polypeptide may possess additional functional domains or an absence of domains. Species variants are polynucleotide sequences that vary from one species to another. The resulting polypeptides generally will have significant amino acid identity relative to each other. A polymorphic variant is a variation in the polynucleotide sequence of a particular gene between individuals of a given species. Polymorphic variants also may encompass “single nucleotide polymorphisms” (SNPs) in which the polynucleotide sequence varies by one base. The presence of SNPs may be indicative of, for example, a certain population, a disease state, or a propensity for a disease state.

The Invention

[0072] The invention is based on the discovery of new human full-length expressed genetic markers (FLEXGEM), the polynucleotides encoding FLEXGEM, and the use of these compositions for the diagnosis, treatment, or prevention of developmental, cell proliferative, and immunological disorders.

[0073] Table 1 lists the Incyte clones used to assemble full length nucleotide sequences encoding FLEXGEM. Columns 1 and 2 show the sequence identification numbers (SEQ ID NOs) of the polypeptide and nucleotide sequences, respectively. Column 3 shows the clone IDs of the Incyte clones in which nucleic acids encoding each FLEXGEM were identified, and column 4 shows the cDNA libraries from which these clones were isolated. Column 5 shows Incyte clones and their corresponding cDNA libraries. Clones for which cDNA libraries are not indicated were derived from pooled cDNA libraries. The clones in column 5 were used to assemble the consensus nucleotide sequence of each FLEXGEM and are useful as fragments in hybridization technologies.

[0074] The columns of Table 2 show various properties of each of the polypeptides of the invention: column 1 references the SEQ ID NO; column 2 shows the number of amino acid residues in each polypeptide; column 3 shows potential phosphorylation sites, and column 4 shows potential glycosylation sites.

[0075] The columns of Table 3 show the tissue-specificity and diseases, disorders, or conditions associated with nucleotide sequences encoding FLEXGEM. The first column of Table 3 lists the nucleotide SEQ ID NOs. Column 2 lists fragments of the nucleotide sequences of column 1. These fragments are useful, for example, in hybridization or amplification technologies to identify SEQ ID NO:21-40 and to distinguish between SEQ ID NO:21-40 and related polynucleotide sequences. The polypeptides encoded by the indicated fragments of SEQ ID NO:21, SEQ ID NO:23-34, 36, 37, and SEQ ID NO:39-40 are useful, for example, as immunogenic peptides. Column 3 lists tissue categories which express FLEXGEM as a fraction of total tissues expressing FLEXGEM. Column 4 lists diseases, disorders, or conditions associated with those tissues expressing FLEXGEM as a fraction of total tissues expressing FLEXGEM. Column 5 lists the vectors used to subclone each cDNA library.

[0076] The columns of Table 4 show descriptions of the tissues used to construct the cDNA libraries from which cDNA clones encoding FLEXGEM were isolated. Column 1 references the nucleotide SEQ ID NOs, column 2 shows the cDNA libraries from which these clones were isolated, and column 3 shows the tissue origins and other descriptive information relevant to the cDNA libraries in column 2.

[0077] The invention incorporates the nucleic acid sequences disclosed in the Sequence Listing and the use of these sequences in the diagnosis and treatment of disease states characterized by altered expression of FLEXGEM genes or defects in FLEXGEM protein function. The invention further utilizes these sequences in hybridization and amplification technologies, and in particular, in technologies which comprehensively assess gene expression patterns correlated with specific cells or tissues and their responses in vivo or in vitro to pharmaceutical agents, toxins, and other treatments. In this manner, the sequences of the present invention are used to develop a transcript image for a particular cell or tissue.

[0078] The invention also encompasses polynucleotides which encode FLEXGEM. In a particular embodiment, the invention encompasses a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID NO:21-40, which encodes FLEXGEM.

[0079] It will be appreciated by those skilled in the art that as a result of the degeneracy of the genetic code, a multitude of polynucleotide sequences encoding FLEXGEM, some bearing minimal similarity to the polynucleotide sequences of any known and naturally occurring gene, may be produced. Thus, the invention contemplates each and every possible variation of polynucleotide sequence that could be made by selecting combinations based on possible codon choices. These combinations are made in accordance with the standard triplet genetic code as applied to the polynucleotide sequence of naturally occurring FLEXGEM, and all such variations are to be considered as being specifically disclosed.

[0080] Although nucleotide sequences which encode FLEXGEM and its variants are preferably capable of hybridizing to the nucleotide sequence of the naturally occurring FLEXGEM under appropriately selected conditions of stringency, it may be advantageous to produce nucleotide sequences encoding FLEXGEM or its derivatives possessing a substantially different codon usage, e.g., inclusion of non-naturally occurring codons. Codons may be selected to increase the rate at which expression of the peptide occurs in a particular prokaryotic or eukaryotic host in accordance with the frequency with which particular codons are utilized by the host. Other reasons for substantially altering the nucleotide sequence encoding FLEXGEM and its derivatives without altering the encoded amino acid sequences include the production of RNA transcripts having more desirable properties, such as a greater half-life, than transcripts produced from the naturally occurring sequence.

[0081] The invention also encompasses production of DNA sequences which encode FLEXGEM and FLEXGEM derivatives, or fragments thereof, entirely by synthetic chemistry. After production, the synthetic sequence may be inserted into any of the many available expression vectors and cell systems using reagents well known in the art. Moreover, synthetic chemistry may be used to introduce mutations into a sequence encoding FLEXGEM or any fragment thereof.

Hybridization

[0082] Also encompassed by the invention are polynucleotide sequences that are capable of hybridizing to the claimed polynucleotide sequences, and, in particular, to those shown in SEQ ID NO:21-40 and fragments thereof under various conditions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399-407; Kimmel, A. R. (1987) Methods Enzymol. 152:507-511.) For example, stringent salt concentration will ordinarily be less than about 750 mM NaCl and 75 mM trisodium citrate, preferably less than about 500 mM NaCl and 50 mM trisodium citrate, and most preferably less than about 250 mM NaCl and 25 mM trisodium citrate. Low stringency hybridization can be obtained in the absence of organic solvent, e.g., formamide, while high stringency hybridization can be obtained in the presence of at least about 35% formamide, and most preferably at least about 50% formamide. Stringent temperature conditions will ordinarily include temperatures of at least about 30° C., more preferably of at least about 37° C., and most preferably of at least about 42° C. Varying additional parameters, such as hybridization time, the concentration of detergent, e.g., sodium dodecyl sulfate (SDS), and the inclusion or exclusion of carrier DNA, are well known to those skilled in the art. Various levels of stringency are accomplished by combining these various conditions as needed. In a preferred embodiment, hybridization will occur at 30° C. in 750 mM NaCl, 75 mM trisodium citrate, and 1% SDS. In a more preferred embodiment, hybridization will occur at 37° C. in 500 mM NaCl, 50 mM trisodium citrate, 1% SDS, 35% formamide, and 100 μg/ml denatured salmon sperm DNA (ssDNA). In a most preferred embodiment, hybridization will occur at 42° C. in 250 mM NaCl, 25 mM trisodium citrate, 1% SDS, 50% formamide, and 200,μg/ml ssDNA. Useful variations on these conditions will be readily apparent to those skilled in the art.

[0083] The washing steps which follow hybridization can also vary in stringency. Wash stringency conditions can be defined by salt concentration and by temperature. As above, wash stringency can be increased by decreasing salt concentration or by increasing temperature. For example, stringent salt concentration for the wash steps will preferably be less than about 30 mM NaCl and 3 mM trisodium citrate, and most preferably less than about 15 mM NaCl and 1.5 mM trisodium citrate. Stringent temperature conditions for the wash steps will ordinarily include temperature of at least about 25° C., more preferably of at least about 42° C., and most preferably of at least about 68° C. In a preferred embodiment, wash steps will occur at 25° C. in 30 mM NaCl, 3 mM trisodium citrate, and 0.1% SDS. In a more preferred embodiment, wash steps will occur at 42° C. in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. In a most preferred embodiment, wash steps will occur at 68° C. in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. Additional variations on these conditions will be readily apparent to those skilled in the art.

cDNA Sequencing

[0084] Methods for DNA sequencing are well known in the art and may be used to practice any of the embodiments of the invention. The methods may employ such enzymes as the Klenow fragment of DNA polymerase I, SEQUENASE (US Biochemical, Cleveland Ohio), Taq polymerase (Perkin-Elmer), thermostable T7 polymerase (Amersham Pharmacia Biotech, Piscataway N.J.), or combinations of polymerases and proofreading exonucleases such as those found in the ELONGASE amplification system (Life Technologies, Gaithersburg Md.). Preferably, sequence preparation is automated with machines such as the MICROLAB 2200 liquid transfer system (Hamilton, Reno Nev.), PTC200 thermal cycler (MJ Research, Watertown Mass.) and ABI CATALYST 800 thermal cycler (Perkin-Elmer). Sequencing is then carried out using either the ABI 373 or 377 DNA sequencing system (Perkin-Elmer), the MEGABACE 1000 DNA sequencing system (Molecular Dynamics, Sunnyvale Calif.), or other systems known in the art. The resulting sequences are analyzed using a variety of algorithms which are well known in the art. (See, e.g., Ausubel, F. M. (1997) Short Protocols in Molecular Biology, John Wiley & Sons, New York N.Y., unit 7.7; Meyers, R. A. (1995) Molecular Biology and Biotechnology, Wiley VCH, New York N.Y., pp. 856-853.)

[0085] The nucleic acid sequences encoding FLEXGEM may be extended utilizing a partial nucleotide sequence and employing various PCR-based methods known in the art to detect upstream sequences, such as promoters and regulatory elements. For example, one method which may be employed, restriction-site PCR, uses universal and nested primers to amplify unknown sequence from genomic DNA within a cloning vector. (See, e.g., Sarkar, G. (1993) PCR Methods Applic. 2:318-322.) Another method, inverse PCR, uses primers that extend in divergent directions to amplify unknown sequence from a circularized template. The template is derived from restriction fragments comprising a known genomic locus and surrounding sequences. (See, e.g., Triglia, T. et al. (1988) Nucleic Acids Res. 16:8186.) A third method, capture PCR, involves PCR amplification of DNA fragments adjacent to known sequences in human and yeast artificial chromosome DNA. (See, e.g., Lagerstrom, M. et al. (1991) PCR Methods Applic. 1:111-119.) In this method, multiple restriction enzyme digestions and ligations may be used to insert an engineered double-stranded sequence into a region of unknown sequence before performing PCR. Other methods which may be used to retrieve unknown sequences are known in the art. (See, e.g., Parker, J. D. et al. (1991) Nucleic Acids Res. 19:3055-306). Additionally, one may use PCR, nested primers, and PROMOTERFINDER libraries (Clontech, Palo Alto Calif.) to walk genomic DNA. This procedure avoids the need to screen libraries and is useful in finding intron/exon junctions. For all PCR-based methods, primers may be designed using commercially available software, such as OLIGO 4.06 Primer Analysis software (National Biosciences, Plymouth Minn.) or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the template at temperatures of about 68° C. to 72° C.

[0086] When screening for full-length cDNAs, it is preferable to use libraries that have been size-selected to include larger cDNAs. In addition, random-primed libraries, which often include sequences containing the 5′ regions of genes, are preferable for situations in which an oligo d(T) library does not yield a full-length cDNA. Genomic libraries may be useful for extension of sequence into 5′ non-transcribed regulatory regions.

[0087] Capillary electrophoresis systems which are commercially available may be used to analyze the size or confirm the nucleotide sequence of sequencing or PCR products. In particular, capillary sequencing may employ flowable polymers for electrophoretic separation, four different nucleotide-specific, laser-stimulated fluorescent dyes, and a charge coupled device camera for detection of the emitted wavelengths. Output/light intensity may be converted to electrical signal using appropriate software (e.g., GENOTYPER and SEQUENCE NAVIGATOR, Perkin-Elmer), and the entire process from loading of samples to computer analysis and electronic data display may be computer controlled. Capillary electrophoresis is especially preferable for sequencing small DNA fragments which may be present in limited amounts in a particular sample.

cDNA Expression

[0088] In another embodiment of the invention, polynucleotide sequences or fragments thereof which encode FLEXGEM may be cloned in recombinant DNA molecules that direct expression of FLEXGEM, or fragments or functional equivalents thereof, in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences which encode substantially the same or a functionally equivalent amino acid sequence may be produced and used to express FLEXGEM.

[0089] The nucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter FLEXGEM-encoding sequences for a variety of purposes including, but not limited to, modification of the cloning, processing, and/or expression of the gene product. DNA shuffling by random fragmentation and PCR reassembly of gene fragments and synthetic oligonucleotides may be used to engineer the nucleotide sequences. For example, oligonucleotide-mediated site-directed mutagenesis may be used to introduce mutations that create new restriction sites, alter glycosylation patterns, change codon preference, produce splice variants, and so forth.

[0090] In another embodiment, sequences encoding FLEXGEM may be synthesized, in whole or in part, using chemical methods well known in the art. (See, e.g., Caruthers, M. H. et al. (1980) Nucl. Acids Res. Symp. Ser. 7:215-223, and Horn, T. et al. (1980) Nucl. Acids Res. Symp. Ser. 7:225-232.) Alternatively, FLEXGEM itself or a fragment thereof may be synthesized using chemical methods. For example, peptide synthesis can be performed using various solid-phase techniques. (See, e.g., Roberge, J. Y. et al. (1995) Science 269:202-204.) Automated synthesis may be achieved using the ABI 431A peptide synthesizer (Perkin-Elmer). Additionally, the amino acid sequence of FLEXGEM, or any part thereof, may be altered during direct synthesis and/or combined with sequences from other proteins, or any part thereof, to produce a variant polypeptide.

[0091] The peptide may be substantially purified by preparative high performance liquid chromatography. (See, e.g, Chiez, R. M. and F. Z. Regnier (1990) Methods Enzymol. 182:392-421.) The composition of the synthetic peptides may be confirmed by amino acid analysis or by sequencing. (See, e.g., Creighton, T. (1984) Proteins, Structures and Molecular Properties, WH Freeman, New York N.Y.)

[0092] In order to express a biologically active FLEXGEM, the nucleotide sequences encoding FLEXGEM or derivatives thereof may be inserted into an appropriate expression vector, i.e., a vector which contains the necessary elements for transcriptional and translational control of the inserted coding sequence in a suitable host. These elements include regulatory sequences, such as enhancers, constitutive and inducible promoters, and 5′ and 3′ untranslated regions in the vector and in polynucleotide sequences encoding FLEXGEM. Such elements may vary in their strength and specificity. Specific initiation signals may also be used to achieve more efficient translation of sequences encoding FLEXGEM. Such signals include the ATG initiation codon and adjacent sequences, e.g. the Kozak sequence. In cases where sequences encoding FLEXGEM and its initiation codon and upstream regulatory sequences are inserted into the appropriate expression vector, no additional transcriptional or translational control signals may be needed. However, in cases where only coding sequence, or a fragment thereof, is inserted, exogenous translational control signals including an in-frame ATG initiation codon should be provided by the vector. Exogenous translational elements and initiation codons may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers appropriate for the particular host cell system used. (See, e.g., Scharf, D. et al. (1994) Results Probl. Cell Differ. 20:125-162.)

[0093] Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding FLEXGEM and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. (See, e.g., Sambrook, J. et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, Plainview N.Y., ch. 4, 8, and 16-17; Ausubel, F. M. et al. (1995) Current Protocols in Molecular Biology, John Wiley & Sons, New York N.Y., ch. 9, 13, and 16.)

[0094] A variety of expression vector/host systems may be utilized to contain and express sequences encoding FLEXGEM. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus); plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV, or tobacco mosaic virus,TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems. The invention is not limited by the host cell employed.

[0095] In bacterial systems, a number of cloning and expression vectors may be selected depending upon the use intended for polynucleotide sequences encoding FLEXGEM. For example, routine cloning, subcloning, and propagation of polynucleotide sequences encoding FLEXGEM can be achieved using a multifunctional E. coli vector such as PBLUESCRIPT (Stratagene, La Jolla Calif.) or pSPORT1 plasmid (Life Technologies). Ligation of sequences encoding FLEXGEM into the vector's multiple cloning site disrupts the lacZ gene, allowing a calorimetric screening procedure for identification of transformed bacteria containing recombinant molecules. In addition, these vectors may be useful for in vitro transcription, dideoxy sequencing, single strand rescue with helper phage, and creation of nested deletions in the cloned sequence. (See, e.g., Van Heeke, G. and S. M. Schuster (1989) J. Biol. Chem. 264:5503-5509.) When large quantities of FLEXGEM are needed, e.g. for the production of antibodies, vectors which direct high level expression of FLEXGEM may be used. For example, vectors containing the strong, inducible T5 or T7 bacteriophage promoter may be used.

[0096] Yeast expression systems may be used for production of FLEXGEM. A number of vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PGH promoters, may be used in the yeast Saccharomyces cerevisiae or Pichia pastoris. In addition, such vectors direct either the secretion or intracellular retention of expressed proteins and enable integration of foreign sequences into the host genome for stable propagation. (See, e.g., Ausubel, 1995, supra; Grant et al. (1987) Methods Enzymol. 153:516-54; and Scorer, C. A. et al. (1994) Bio/Technology 12:181-184.) Plant systems may also be used for expression of FLEXGEM. Transcription of sequences encoding FLEXGEM may be driven viral promoters, e.g., the 35S and 19S promoters of CaMV used alone or in combination with the omega leader sequence from TMV (Takamatsu, N. (1987) EMBO J. 6:307-311). Alternatively, plant promoters such as the small subunit of RUBISCO or heat shock promoters may be used. (See, e.g., Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Broglie, R. et al. (1984) Science 224:838-843; and Winter, J. et al. (1991) Results Probl. Cell Differ. 17:85-105.) These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. (See, e.g., The McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York N.Y., pp. 191-196.)

[0097] In mammalian cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, sequences encoding FLEXGEM may be ligated into an adenovirus transcription/translation complex consisting of the late promoter and tripartite leader sequence. Insertion in a non-essential E1 or E3 region of the viral genome may be used to obtain infective virus which expresses FLEXGEM in host cells. (See, e.g., Logan, J. and T. Shenk (1984) Proc. Natl. Acad. Sci. 81:3655-3659.) In addition, transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer, may be used to increase expression in mammalian host cells. SV40 or EBV-based vectors may also be used for high-level protein expression.

[0098] Human artificial chromosomes (HACs) may also be employed to deliver larger fragments of DNA than can be contained in and expressed from a plasmid. HACs of about 6 kb to 10 Mb are constructed and delivered via conventional delivery methods (liposomes, polycationic amino polymers, or vesicles) for therapeutic purposes. (See, e.g., Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355.)

[0099] For long term production of recombinant proteins in mammalian systems, stable expression of FLEXGEM in cell lines is preferred. For example, sequences encoding FLEXGEM can be transformed into cell lines using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. Following the introduction of the vector, cells may be allowed to grow for about 1 to 2 days in enriched media before being switched to selective media. The purpose of the selectable marker is to confer resistance to a selective agent, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate to the cell type.

[0100] Any number of selection systems may be used to recover transformed cell lines. These include, but are not limited to, the herpes simplex virus thymidine kinase and adenine phosphoribosyltransferase genes, for use in tk- or apr- cells, respectively. (See, e.g., Wigler, M. et al. (1977) Cell 11:223-232; Lowy, I. et al. (1980) Cell 22:817-823.) Also, antimetabolite, antibiotic, or herbicide resistance can be used as the basis for selection. For example, dhfr confers resistance to methotrexate; neo confers resistance to the aminoglycosides neomycin and G-418; and als or pat confer resistance to chlorsulfuron and phosphinotricin acetyltransferase, respectively. (See, e.g., Wigler, M. et al. (1980) Proc. Natl. Acad. Sci. 77:3567-3570; Colbere-Garapin, F. et al. (1981) J. Mol. Biol. 150:1-14.) Additional selectable genes have been described, e.g., trpB and hisD, which alter cellular requirements for metabolites. (See, e.g., Hartman, S. C. and R. C. Mulligan (1988) Proc. Natl. Acad. Sci. 85:8047-8051.) Visible markers, e.g., anthocyanins, green fluorescent proteins (GFP; Clontech), β glucuronidase and its substrate β-glucuronide, or luciferase and its substrate luciferin may be used. These markers can be used not only to identify transformants, but also to quantify the amount of transient or stable protein expression attributable to a specific vector system. (See, e.g., Rhodes, C. A. (1995) Methods Mol. Biol. 55:121-131.)

[0101] Although the presence/absence of marker gene expression suggests that the gene of interest is also present, the presence and expression of the gene may need to be confirmed. For example, if the sequence encoding FLEXGEM is inserted within a marker gene sequence, transformed cells containing sequences encoding FLEXGEM can be identified by the absence of marker gene function. Alternatively, a marker gene can be placed in tandem with a sequence encoding FLEXGEM under the control of a single promoter. Expression of the marker gene in response to induction or selection usually indicates expression of the tandem gene as well.

[0102] In general, host cells that contain the nucleic acid sequence encoding FLEXGEM and that express FLEXGEM may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA-DNA or DNA-RNA hybridizations, PCR amplification, and protein bioassay or immunoassay techniques which include membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or protein sequences.

[0103] Immunological methods for detecting and measuring the expression of FLEXGEM using either specific polyclonal or monoclonal antibodies are known in the art. Examples of such techniques include enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and fluorescence activated cell sorting (FACS). A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes on FLEXGEM is preferred, but a competitive binding assay may be employed. These and other assays are well known in the art. (See, e.g., Hampton, R. et al. (1990) Serological Methods, a Laboratory Manual, APS Press, St Paul Minn., Sect. IV; Coligan, J. E. et al. (1997) Current Protocols in Immunology, Greene Pub. Associates and Wiley-Interscience, New York N.Y.; and Pound, J. D. (1998) Immunochemical Protocols, Humana Press, Totowa N.J.).

[0104] A wide variety of labels and conjugation techniques are known by those skilled in the art and may be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides encoding FLEXGEM include oligolabeling, nick translation, end-labeling, or PCR amplification using a labeled nucleotide. Alternatively, the sequences encoding FLEXGEM, or any fragments thereof, may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by addition of an appropriate RNA polymerase such as T7, T3, or SP6 and labeled nucleotides. These procedures may be conducted using a variety of commercially available kits, such as those provided by Amersham Pharmacia Biotech, Promega (Madison Wis.), and US Biochemical. Suitable reporter molecules or labels which may be used for ease of detection include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents, as well as substrates, cofactors, inhibitors, magnetic particles, and the like.

[0105] Host cells transformed with nucleotide sequences encoding FLEXGEM may be cultured under conditions suitable for the expression and recovery of the protein from cell culture. The protein produced by a transformed cell may be secreted or retained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides which encode FLEXGEM may be designed to contain signal sequences which direct secretion of FLEXGEM through a prokaryotic or eukaryotic cell membrane.

[0106] In addition, a host cell strain may be chosen for its ability to modulate expression of the inserted sequences or to process the expressed protein in the desired fashion. Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing which cleaves a “prepro” form of the protein may also be used to specify protein targeting, folding, and/or activity. Different host cells which have specific cellular machinery and characteristic mechanisms for post-translational activities (e.g., CHO, HeLa, MDCK, HEK293, and WI38) are available from the American Type Culture Collection (ATCC, Bethesda MD) and may be chosen to ensure the correct modification and processing of the foreign protein.

[0107] In another embodiment of the invention, natural, modified, or recombinant nucleic acid sequences encoding FLEXGEM may be ligated to a heterologous sequence resulting in translation of a fusion protein in any of the aforementioned host systems. For example, a chimeric FLEXGEM protein containing a heterologous moiety that can be recognized by a commercially available antibody may facilitate the screening of peptide libraries for inhibitors of FLEXGEM activity. Heterologous protein and peptide moieties may also facilitate purification of fusion proteins using commercially available affinity matrices. Such moieties include, but are not limited to, glutathione S-transferase (GST), maltose binding protein (MBP), thioredoxin (Trx), calmodulin binding peptide (CBP), 6-His, FLAG, c-myc, and hemagglutinin (HA). GST, MBP, Trx, CBP, and 6-His enable purification of their cognate fusion proteins on immobilized glutathione, maltose, phenylarsine oxide, calmodulin, and metal-chelate resins, respectively. FLAG, c-myc, and hemagglutinin (HA) enable immunoaffinity purification of fusion proteins using commercially available monoclonal and polyclonal antibodies that specifically recognize these epitope tags. A fusion protein may also be engineered to contain a proteolytic cleavage site located between the FLEXGEM encoding sequence and the heterologous protein sequence, so that FLEXGEM may be cleaved away from the heterologous moiety following purification. Methods for fusion protein expression and purification are discussed in Ausubel (1995, supra, ch 10). A variety of commercially available kits may also be used to facilitate expression and purification of fusion proteins.

[0108] In a further embodiment of the invention, synthesis of radiolabeled FLEXGEM may be achieved in vitro using the TNT rabbit reticulocyte lysate or wheat germ extract systems (Promega). These systems couple transcription and translation of protein-coding sequences operably associated with the T7, T3, or SP6 promoters. Translation takes place in the presence of a radiolabeled amino acid precursor, preferably ³⁵S-methionine.

[0109] Fragments of FLEXGEM may be produced not only by recombinant production, but also by direct peptide synthesis using solid-phase techniques. (See, e.g., Creighton, supra, pp. 55-60.) Protein synthesis may be performed by manual techniques or by automation. Automated synthesis may be achieved, for example, using the ABI 431A peptide synthesizer (Perkin-Elmer). Various fragments of FLEXGEM may be synthesized separately and then combined to produce the full length molecule.

Screening Assays

[0110] FLEXGEM encoded by polynucleotides of the present invention may be used to screen for molecules that bind to or are bound by the encoded polypeptides. The binding of the polypeptide and the molecule may activate (agonist), increase, inhibit (antagonist), or decrease activity of the polypeptide or the bound molecule. Examples of such molecules include antibodies, oligonucleotides, proteins (e.g., receptors), or small molecules.

[0111] Preferably, the molecule is closely related to the natural ligand of the polypeptide, e.g., a ligand or fragment thereof, a natural substrate, or a structural or functional mimetic. (See, Coligan et al., (1991) Current Protocols in Immunology 1(2): Chapter 5.) Similarly, the molecule can be closely related to the natural receptor to which the polypeptide binds, or to at least a fragment of the receptor, e.g., the active site. In either case, the molecule can be rationally designed using known techniques. Preferably, the screening for these molecules involves producing appropriate cells which express the polypeptide, either as a secreted protein or on the cell membrane. Preferred cells include cells from mammals, yeast, Drosophila, or E. coli. Cells expressing the polypeptide or cell membrane fractions which contain the expressed polypeptide are then contacted with a test compound and binding, stimulation, or inhibition of activity of either the polypeptide or the molecule is analyzed.

[0112] An assay may simply test binding of a candidate compound to the polypeptide, wherein binding is detected by a fluorophore, radioisotope, enzyme conjugate, or other detectable label. Alternatively, the assay may assess binding in the presence of a labeled competitor.

[0113] Additionally, the assay can be carried out using cell-free preparations, polypeptide/molecule affixed to a solid support, chemical libraries, or natural product mixtures. The assay may also simply comprise the steps of mixing a candidate compound with a solution containing a polypeptide, measuring polypeptide/molecule activity or binding, and comparing the polypeptide/molecule activity or binding to a standard.

[0114] Preferably, an ELISA assay using, e.g., a monoclonal or polyclonal antibody, can measure polypeptide level in a sample. The antibody can measure polypeptide level by either binding, directly or indirectly, to the polypeptide or by competing with the polypeptide for a substrate.

[0115] All of the above assays can be used in a diagnostic or prognostic context. The molecules discovered using these assays can be used to treat disease or to bring about a particular result in a patient (e.g., blood vessel growth) by activating or inhibiting the polypeptide/molecule. Moreover, the assays can discover agents which may inhibit or enhance the production of the polypeptide from suitably manipulated cells or tissues.

Transcript Imaging

[0116] Another embodiment relates to the use of polynucleotide sequences encoding FLEXGEM to develop a transcript image of a tissue or cell type. A transcript image is the collective pattern of gene expression by a particular tissue or cell type under given conditions and at a given time. This pattern of gene expression is defined by the number of expressed genes and their abundance. Thus the polynucleotide sequences of the present invention may be used to develop a transcript image of a tissue or cell type by hybridizing, preferably in a microarray format, the polynucleotide sequences of the present invention to the totality of transcripts or reverse transcripts of a tissue or cell type. The resultant transcript image would provide a profile of FLEXGEM gene activity.

[0117] Transcript images which profile FLEXGEM gene expression may be generated using transcripts isolated from tissues, cell lines, biopsies, or other biological samples. The transcript image may thus reflect FLEXGEM gene expression in vivo, as in the case of a tissue or biopsy sample, or in vitro, as in the case of a cell line. Transcript images may be used to profile FLEXGEM gene expression in distinct tissue types. This process can be used to determine FLEXGEM gene activity in a particular tissue type relative to this activity in a different tissue type. Transcript images may be used to generate a profile of FLEXGEM gene expression characteristic of diseased tissue. Transcript images of tissues before and after treatment may be used for diagnostic purposes, to monitor the progression of disease, and to monitor the efficacy of drug treatments for diseases which affect the activity of genes encoding FLEXGEM.

[0118] Transcript images which profile FLEXGEM gene expression may also be used in conjunction with in vitro model systems and preclinical evaluation of pharmaceuticals. Transcript images of cell lines can be used to assess FLEXGEM activity and/or to identify cell lines that lack or misregulate this activity. Such cell lines may then be treated with pharmaceutical agents, and a transcript image following treatment may indicate the efficacy of these agents in restoring desired levels of this activity. A similar approach may be used to assess the toxicity of pharmaceutical agents as reflected by undesirable changes in FLEXGEM activity. Candidate pharmaceutical agents may be evaluated by comparing their associated transcript images with those of pharmaceutical agents of known effectiveness.

Therapeutics

[0119] The expression of FLEXGEM is closely associated with cell proliferation and the immune response. Therefore, FLEXGEM appears to play a role in developmental, cell proliferative, and immunological disorders. In the treatment of disorders associated with increased FLEXGEM expression or activity, it is desirable to decrease the expression or activity of FLEXGEM. In the treatment of disorders associated with decreased FLEXGEM expression or activity, it is desirable to increase the expression or activity of FLEXGEM.

[0120] Therefore, in one embodiment, FLEXGEM or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of FLEXGEM. Examples of such disorders include, but are not limited to, a developmental disorder such as renal tubular acidosis, anemia, Cushing's syndrome, achondroplastic dwarfism, Duchenne and Becker muscular dystrophy, epilepsy, gonadal dysgenesis, WAGR syndrome (Wilms' tumor, aniridia, genitourinary abnormalities, and mental retardation), Smith-Magenis syndrome, myelodysplastic syndrome, hereditary mucoepithelial dysplasia, hereditary keratodermas, hereditary neuropathies such as Charcot-Marie-Tooth disease and neurofibromatosis, hypothyroidism, hydrocephalus, a seizure disorder such as Syndenham's chorea and cerebral palsy, spina bifida, anencephaly, craniorachischisis, congenital glaucoma, cataract, and sensorineural hearing loss; a cell proliferative disorder such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, a cancer of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus; and an immunological disorder such as inflammation, actinic keratosis, acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, arteriosclerosis, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, bronchitis, bursitis, cholecystitis, cirrhosis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, paroxysmal nocturnal hemoglobinuria, hepatitis, hypereosinophilia, irritable bowel syndrome, episodic lymphopenia with lymphocytotoxins, mixed connective tissue disease (MCTD), multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, myelofibrosis, osteoarthritis, osteoporosis, pancreatitis, polycythemia vera, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjögren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, primary thrombocythemia, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, trauma, and hematopoietic cancer including lymphoma, leukemia, and myeloma.

[0121] In another embodiment, a vector capable of expressing FLEXGEM or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of FLEXGEM including, but not limited to, those described above.

[0122] In a further embodiment, a pharmaceutical composition comprising a substantially purified FLEXGEM in conjunction with a suitable pharmaceutical carrier may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of FLEXGEM including, but not limited to, those provided above.

[0123] In still another embodiment, an agonist which modulates the activity of FLEXGEM may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of FLEXGEM including, but not limited to, those listed above.

[0124] In a further embodiment, an antagonist of FLEXGEM may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of FLEXGEM. Examples of such disorders include, but are not limited to, those described above. In one aspect, an antibody which specifically binds FLEXGEM may be used directly as an antagonist or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissue which express FLEXGEM.

[0125] In an additional embodiment, a vector expressing the complement of the polynucleotide encoding FLEXGEM may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of FLEXGEM including, but not limited to, those described above.

[0126] In other embodiments, any of the proteins, antagonists, antibodies, agonists, complementary sequences, or vectors of the invention may be administered in combination with other appropriate therapeutic agents. Selection of the appropriate agents for use in combination therapy may be made by one of ordinary skill in the art, according to conventional pharmaceutical principles. The combination of therapeutic agents may act synergistically to effect the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent, thus reducing the potential for adverse side effects.

[0127] An antagonist of FLEXGEM may be produced using methods which are generally known in the art. In particular, purified FLEXGEM may be used to produce antibodies or to screen libraries of pharmaceutical agents to identify those which specifically bind FLEXGEM. Antibodies to FLEXGEM may also be generated using methods that are well known in the art. Such antibodies may include, but are not limited to, polyclonal, monoclonal, chimeric, and single chain antibodies, Fab fragments, and fragments produced by a Fab expression library. Neutralizing antibodies (i.e., those which inhibit dimer formation) are especially preferred for therapeutic use.

[0128] For the production of antibodies, various hosts including goats, rabbits, rats, mice, humans, and others may be immunized by injection with FLEXGEM or with any fragment or oligopeptide thereof which has immunogenic properties. Depending on the host species, various adjuvants may be used to increase immunological response. Such adjuvants include, but are not limited to, Freund's, mineral gels such as aluminum hydroxide, and surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, KLH, and dinitrophenol. Among adjuvants used in humans, BCG (bacilli Calmette-Guerin) and Corynebacterium parvum are especially preferable.

[0129] It is preferred that the oligopeptides, peptides, or fragments used to induce antibodies to FLEXGEM have an amino acid sequence consisting of at least about 5 amino acids, and, more preferably, of at least about 10 amino acids. It is also preferable that these oligopeptides, peptides, or fragments are identical to a portion of the amino acid sequence of the natural protein and contain the entire amino acid sequence of a small, naturally occurring molecule. Short stretches of FLEXGEM amino acids may be fused with those of another protein, such as KLH, and antibodies to the chimeric molecule may be produced.

[0130] Monoclonal antibodies to FLEXGEM may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the EBV-hybridoma technique. (See, e.g., Kohler, G. et al. (1975) Nature 256:495-497; Kozbor, D. et al. (1985) J. Immunol. Methods 81:31-42; Cote, R. J. et al. (1983) Proc. Natl. Acad. Sci. 80:2026-2030; and Cole, S. P. et al. (1984) Mol. Cell Biol. 62:109-120.)

[0131] In addition, techniques developed for the production of “chimeric antibodies,” such as the splicing of mouse antibody genes to human antibody genes to obtain a molecule with appropriate antigen specificity and biological activity, can be used. (See, e.g., Morrison, S. L. et al. (1984) Proc. Natl. Acad. Sci. 81:6851-6855; Neuberger, M. S. et al. (1984) Nature 312:604-608; and Takeda, S. et al. (1985) Nature 314:452-454.) Alternatively, techniques described for the production of single chain antibodies may be adapted, using methods known in the art, to produce FLEXGEM-specific single chain antibodies. Antibodies with related specificity, but of distinct idiotypic composition, may be generated by chain shuffling from random combinatorial immunoglobulin libraries. (See, e.g., Burton D. R. (1991) Proc. Natl. Acad. Sci. 88:10134-10137.)

[0132] Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening immunoglobulin libraries or panels of highly specific binding reagents as disclosed in the literature. (See, e.g., Orlandi, R. et al. (1989) Proc. Natl. Acad. Sci. 86: 3833-3837; Winter, G. et al. (1991) Nature 349:293-299.)

[0133] Antibody fragments which contain specific binding sites for FLEXGEM may also be generated. For example, such fragments include, but are not limited to, F(ab′)2 fragments produced by pepsin digestion of the antibody molecule and Fab fragments generated by reducing the disulfide bridges of the F(ab′)2 fragments. Alternatively, Fab expression libraries may be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity. (See, e.g., Huse, W. D. et al. (1989) Science 246:1275-1281.)

[0134] Various immunoassays may be used for screening to identify antibodies having the desired specificity. Numerous protocols for competitive binding or immunoradiometric assays using either polyclonal or monoclonal antibodies with established specificities are well known in the art. Such immunoassays typically involve the measurement of complex formation between FLEXGEM and its specific antibody. A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering FLEXGEM epitopes is preferred, but a competitive binding assay may also be employed (Pound, supra).

[0135] Various methods such as Scatchard analysis in conjunction with radioimmunoassay techniques may be used to assess the affinity of antibodies for FLEXGEM. Affinity is expressed as an association constant, K_(a), which is defined as the molar concentration of FLEXGEM-antibody complex divided by the molar concentrations of free antigen and free antibody under equilibrium conditions. The K_(a) determined for a preparation of polyclonal antibodies, which are heterogeneous in their affinities for multiple FLEXGEM epitopes, represents the average affinity, or avidity, of the antibodies for FLEXGEM. The K_(a) determined for a preparation of monoclonal antibodies, which are monospecific for a particular FLEXGEM epitope, represents a true measure of affinity. High-affinity antibody preparations with K_(a) ranging from about 10⁹ to 10¹² L/mole are preferred for use in immunoassays in which the FLEXGEM-antibody complex must withstand rigorous manipulations. Low-affinity antibody preparations with K_(a) ranging from about 10⁶ to 10⁷ L/mole are preferred for use in immunopurification and similar procedures which ultimately require dissociation of FLEXGEM, preferably in active form, from the antibody (Catty, D. (1988) Antibodies, Volume I: A Practical Approach, IRL Press, Washington, D.C.; Liddell, J. E. and Cryer, A. (1991) A Practical Guide to Monoclonal Antibodies, John Wiley & Sons, New York N.Y.).

[0136] The titer and avidity of polyclonal antibody preparations may be further evaluated to determine the quality and suitability of such preparations for certain downstream applications. For example, a polyclonal antibody preparation containing at least 1-2 mg specific antibody/ml, preferably 5-10 mg specific antibody/ml, is preferred for use in procedures requiring precipitation of FLEXGEM-antibody complexes. Procedures for evaluating antibody specificity, titer, and avidity, and guidelines for antibody quality and usage in various applications, are generally available. (See, e.g., Catty, supra, and Coligan et al. supra.)

[0137] In another embodiment of the invention, the polynucleotides encoding FLEXGEM, or any fragment or complement thereof, may be used for therapeutic purposes. In one aspect, the complement of the polynucleotide encoding FLEXGEM may be used in situations in which it would be desirable to block the transcription of the mRNA. In particular, cells may be transformed with sequences complementary to polynucleotides encoding FLEXGEM. Thus, complementary molecules or fragments may be used to modulate FLEXGEM activity, or to achieve regulation of gene function. Such technology is now well known in the art, and sense or antisense oligonucleotides or larger fragments can be designed from various locations along the coding or control regions of sequences encoding FLEXGEM.

[0138] Expression vectors derived from retroviruses, adenoviruses, or herpes or vaccinia viruses, or from various bacterial plasmids, may be used for delivery of nucleotide sequences to the targeted organ, tissue, or cell population. Methods which are well known to those skilled in the art can be used to construct vectors to express nucleic acid sequences complementary to the polynucleotides encoding FLEXGEM. (See, e.g., Sambrook, supra; Ausubel, 1995, supra.)

[0139] Genes encoding FLEXGEM can be turned off by transforming a cell or tissue with expression vectors which express high levels of a polynucleotide, or fragment thereof, encoding FLEXGEM. Such constructs may be used to introduce untranslatable sense or antisense sequences into a cell. Even in the absence of integration into the DNA, such vectors may continue to transcribe RNA molecules until they are disabled by endogenous nucleases. Transient expression may last for a month or more with a non-replicating vector, and may last even longer if appropriate replication elements are part of the vector system.

[0140] As mentioned above, modifications of gene expression can be obtained by designing complementary sequences or antisense molecules (DNA, RNA, or PNA) to the control, 5′, or regulatory regions of the gene encoding FLEXGEM. Oligonucleotides derived from the transcription initiation site, e.g., between about positions −10 and +10 from the start site, are preferred. Similarly, inhibition can be achieved using triple helix base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or regulatory molecules. Recent therapeutic advances using triplex DNA have been described in the literature. (See, e.g., Gee, J. E. et al. (1994) in Huber, B. E. and B. I. Carr, Molecular and Immunologic Approaches, Futura Publishing, Mt. Kisco N.Y., pp. 163-177.) A complementary sequence or antisense molecule may also be designed to block translation of mRNA by preventing the transcript from binding to ribosomes.

[0141] Ribozymes, enzymatic RNA molecules, may also be used to catalyze the specific cleavage of RNA. The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage. For example, engineered hammerhead motif ribozyme molecules may specifically and efficiently catalyze endonucleolytic cleavage of sequences encoding FLEXGEM.

[0142] Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites, including the following sequences: GUA, GUU, and GUC. Once identified, short RNA sequences of between 15 and 20 ribonucleotides, corresponding to the region of the target gene containing the cleavage site, may be evaluated for secondary structural features which may render the oligonucleotide inoperable. The suitability of candidate targets may also be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays.

[0143] Complementary ribonucleic acid molecules and ribozymes of the invention may be prepared by any method known in the art for the synthesis of nucleic acid molecules. These include techniques for chemically synthesizing oligonucleotides such as solid phase phosphoramidite chemical synthesis. Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA sequences encoding FLEXGEM. Such DNA sequences may be incorporated into a wide variety of vectors with suitable RNA polymerase promoters such as T7 or SP6. Alternatively, these cDNA constructs that synthesize complementary RNA, constitutively or inducibly, can be introduced into cell lines, cells, or tissues.

[0144] RNA molecules may be modified to increase intracellular stability and half-life. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5′ and/or 3′ ends of the molecule, or the use of phosphorothioate or 2′ O-methyl rather than phosphodiesterase linkages within the backbone of the molecule. This concept is inherent in the production of PNAs and can be extended in all of these molecules by the inclusion of nontraditional bases such as inosine, queosine, and wybutosine, as well as acetyl-, methyl-, thio-, and similarly modified forms of adenine, cytidine, guanine, thymine, and uridine which are not as easily recognized by endogenous endonucleases.

[0145] Many methods for introducing vectors into cells or tissues are available and equally suitable for use in vivo, in vitro, and ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from the patient and clonally propagated for autologous transplant back into that same patient. Delivery by transfection, by liposome injections, or by polycationic amino polymers may be achieved using methods which are well known in the art. (See, e.g., Goldman, C. K. et al. (1997) Nature Biotechnology 15:462-466.)

[0146] Any of the therapeutic methods described above may be applied to any subject in need of such therapy, including, for example, mammals such as dogs, cats, cows, horses, rabbits, monkeys, and most preferably, humans.

[0147] An additional embodiment of the invention relates to the administration of a pharmaceutical or sterile composition, in conjunction with a pharmaceutically acceptable carrier, for any of the therapeutic effects discussed above. Such pharmaceutical compositions may consist of FLEXGEM, antibodies to FLEXGEM, and mimetics, agonists, antagonists, or inhibitors of FLEXGEM. The compositions may be administered alone or in combination with at least one other agent, such as a stabilizing compound, which may be administered in any sterile, biocompatible pharmaceutical carrier including, but not limited to, saline, buffered saline, dextrose, and water. The compositions may be administered to a patient alone, or in combination with other agents, drugs, or hormones.

[0148] The pharmaceutical compositions utilized in this invention may be administered by any number of routes including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal means.

[0149] In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically-acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Further details on techniques for formulation and administration may be found in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing, Easton Pa.).

[0150] Pharmaceutical compositions for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration. Such carriers enable the pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for ingestion by the patient.

[0151] Pharmaceutical preparations for oral use can be obtained through combining active compounds with solid excipient and processing the resultant mixture of granules (optionally, after grinding) to obtain tablets or dragee cores. Suitable auxiliaries can be added, if desired. Suitable excipients include carbohydrate or protein fillers, such as sugars, including lactose, sucrose, mannitol, and sorbitol; starch from corn, wheat, rice, potato, or other plants; cellulose, such as methyl cellulose, hydroxypropylmethyl-cellulose, or sodium carboxymethylcellulose; gums, including arabic and tragacanth; and proteins, such as gelatin and collagen. If desired, disintegrating or solubilizing agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, and alginic acid or a salt thereof, such as sodium alginate.

[0152] Dragee cores may be used in conjunction with suitable coatings, such as concentrated sugar solutions, which may also contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for product identification or to characterize the quantity of active compound, i.e., dosage.

[0153] Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a coating, such as glycerol or sorbitol. Push-fit capsules can contain active ingredients mixed with fillers or binders, such as lactose or starches, lubricants, such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid, or liquid polyethylene glycol with or without stabilizers.

[0154] Pharmaceutical formulations suitable for parenteral administration may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks' solution, Ringer's solution, or physiologically buffered saline. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate, triglycerides, or liposomes. Non-lipid polycationic amino polymers may also be used for delivery. Optionally, the suspension may also contain suitable stabilizers or agents to increase the solubility of the compounds and allow for the preparation of highly concentrated solutions.

[0155] For topical or nasal administration, penetrants appropriate to the particular barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0156] The pharmaceutical compositions of the present invention may be manufactured in a manner that is known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes.

[0157] The pharmaceutical composition may be provided as a salt and can be formed with many acids, including but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, and succinic acids. Salts tend to be more soluble in aqueous or other protonic solvents than are the corresponding free base forms. In other cases, the preferred preparation may be a lyophilized powder which may contain any or all of the following: 1 mM to 50 mM histidine, 0.1% to 2% sucrose, and 2% to 7% mannitol, at a pH range of 4.5 to 5.5, that is combined with buffer prior to use.

[0158] After pharmaceutical compositions have been prepared, they can be placed in an appropriate container and labeled for treatment of an indicated condition. For administration of FLEXGEM, such labeling would include amount, frequency, and method of administration.

[0159] Pharmaceutical compositions suitable for use in the invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. The determination of an effective dose is well within the capability of those skilled in the art.

[0160] For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays, e.g., of neoplastic cells or in animal models such as mice, rats, rabbits, dogs, or pigs. An animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans.

[0161] A therapeutically effective dose refers to that amount of active ingredient, for example FLEXGEM or fragments thereof, antibodies of FLEXGEM, and agonists, antagonists or inhibitors of FLEXGEM, which ameliorates the symptoms or condition. Therapeutic efficacy and toxicity may be determined by standard pharmaceutical procedures in cell cultures or with experimental animals, such as by calculating the ED₅₀ (the dose therapeutically effective in 50% of the population) or LD₅₀ (the dose lethal to 50% of the population) statistics. The dose ratio of toxic to therapeutic effects is the therapeutic index, which can be expressed as the LD₅/ED₅₀ ratio. Pharmaceutical compositions which exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used to formulate a range of dosage for human use. The dosage contained in such compositions is preferably within a range of circulating concentrations that includes the ED₅₀ with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, the sensitivity of the patient, and the route of administration.

[0162] The exact dosage will be determined by the practitioner, in light of factors related to the subject requiring treatment. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, the general health of the subject, the age, weight, and gender of the subject, time and frequency of administration, drug combination(s), reaction sensitivities, and response to therapy. Long-acting pharmaceutical compositions may be administered every 3 to 4 days, every week, or biweekly depending on the half-life and clearance rate of the particular formulation.

[0163] Normal dosage amounts may vary from about 0.1 μg to 100,000 μg, up to a total dose of about 1 gram, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art. Those skilled in the art will employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc.

Diagnostics

[0164] In another embodiment, antibodies which specifically bind FLEXGEM may be used for the diagnosis of disorders characterized by expression of FLEXGEM, or in assays to monitor patients being treated with FLEXGEM or agonists, antagonists, or inhibitors of FLEXGEM. Antibodies useful for diagnostic purposes may be prepared in the same manner as described above for therapeutics. Diagnostic assays for FLEXGEM include methods which utilize the antibody and a label to detect FLEXGEM in human body fluids or in extracts of cells or tissues. The antibodies may be used with or without modification, and may be labeled by covalent or non-covalent attachment of a reporter molecule. A wide variety of reporter molecules, several of which are described above, are known in the art and may be used.

[0165] A variety of protocols for measuring FLEXGEM, including ELISAs, RIAs, and FACS, are known in the art and provide a basis for diagnosing altered or abnormal levels of FLEXGEM expression. Normal or standard values for FLEXGEM expression are established by combining body fluids or cell extracts taken from normal mammalian subjects, preferably human, with antibody to FLEXGEM under conditions suitable for complex formation. The amount of standard complex formation may be quantitated by various methods, preferably by photometric means. Quantities of FLEXGEM expressed in subject, control, and disease samples from biopsied tissues are compared with the standard values. Deviation between standard and subject values establishes the parameters for diagnosing disease.

[0166] In another embodiment of the invention, the polynucleotides encoding FLEXGEM may be used for diagnostic purposes. The polynucleotides which may be used include oligonucleotide sequences, complementary RNA and DNA molecules, and PNAs. The polynucleotides may be used to detect and quantitate gene expression in biopsied tissues in which expression of FLEXGEM may be correlated with disease. The diagnostic assay may be used to determine absence, presence, and excess expression of FLEXGEM, and to monitor regulation of FLEXGEM levels during therapeutic intervention.

[0167] In one aspect, hybridization with PCR probes which are capable of detecting polynucleotide sequences, including genomic sequences, encoding FLEXGEM or closely related molecules may be used to identify nucleic acid sequences which encode FLEXGEM. The specificity of the probe, whether it is made from a highly specific region, e.g., the 5′ regulatory region, or from a less specific region, e.g., a conserved motif, and the stringency of the hybridization or amplification (maximal, high, intermediate, or low), will determine whether the probe identifies only naturally occurring sequences encoding FLEXGEM, allelic variants, or related sequences.

[0168] Probes may also be used for the detection of related sequences, and should preferably have at least 50% sequence identity to any of the FLEXGEM encoding sequences. The hybridization probes of the subject invention may be DNA or RNA and may be derived from the sequence of SEQ ID NO:21-40 or from genomic sequences including promoters, enhancers, and introns of the FLEXGEM gene.

[0169] Means for producing specific hybridization probes for DNAs encoding FLEXGEM include the cloning of polynucleotide sequences encoding FLEXGEM or FLEXGEM derivatives into vectors for the production of mRNA probes. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by means of the addition of the appropriate RNA polymerases and the appropriate labeled nucleotides. Hybridization probes may be labeled by a variety of reporter groups, for example, by radionuclides such as ³²P or ₃₅S, or by enzymatic labels, such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems, and the like.

[0170] Polynucleotide sequences encoding FLEXGEM may be used for the diagnosis of disorders associated with expression of FLEXGEM. Examples of such disorders include, but are not limited to, a developmental disorder such as renal tubular acidosis, anemia, Cushing's syndrome, achondroplastic dwarfism, Duchenne and Becker muscular dystrophy, epilepsy, gonadal dysgenesis, WAGR syndrome (Wilms' tumor, aniridia, genitourinary abnormalities, and mental retardation), Smith-Magenis syndrome, myelodysplastic syndrome, hereditary mucoepithelial dysplasia, hereditary keratodermas, hereditary neuropathies such as Charcot-Marie-Tooth disease and neurofibromatosis, hypothyroidism, hydrocephalus, a seizure disorder such as Syndenham's chorea and cerebral palsy, spina bifida, anencephaly, craniorachischisis, congenital glaucoma, cataract, and sensorineural hearing loss; a cell proliferative disorder such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, a cancer of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus; and an immunological disorder such as inflammation, actinic keratosis, acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, arteriosclerosis, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, bronchitis, bursitis, cholecystitis, cirrhosis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, paroxysmal nocturnal hemoglobinuria, hepatitis, hypereosinophilia, irritable bowel syndrome, episodic lymphopenia with lymphocytotoxins, mixed connective tissue disease (MCTD), multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, myelofibrosis, osteoarthritis, osteoporosis, pancreatitis, polycythemia vera, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjögren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, primary thrombocythemia, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, trauma, and hematopoietic cancer including lymphoma, leukemia, and myeloma. The polynucleotide sequences encoding FLEXGEM may be used in Southern or northern analysis, dot blot, or other membrane-based technologies; in PCR technologies; in dipstick, pin, and multiformat ELISA-like assays; and in microarrays utilizing fluids or tissues from patients to detect altered FLEXGEM expression. Such qualitative or quantitative methods are well known in the art.

[0171] In a particular aspect, the nucleotide sequences encoding FLEXGEM may be useful in assays that detect the presence of associated disorders, particularly those mentioned above. The nucleotide sequences encoding FLEXGEM may be labeled by standard methods and added to a fluid or tissue sample from a patient under conditions suitable for the formation of hybridization complexes. After a suitable incubation period, the sample is washed and the signal is quantitated and compared with a standard value. If the amount of signal in the patient sample is significantly altered in comparison to a control sample then the presence of altered levels of nucleotide sequences encoding FLEXGEM in the sample indicates the presence of the associated disorder. Such assays may also be used to evaluate the efficacy of a particular therapeutic treatment regimen in animal studies, in clinical trials, or to monitor the treatment of an individual patient.

[0172] In order to provide a basis for the diagnosis of a disorder associated with expression of FLEXGEM, a normal or standard profile for expression is established. This may be accomplished by combining body fluids or cell extracts taken from normal subjects, either animal or human, with a sequence, or a fragment thereof, encoding FLEXGEM, under conditions suitable for hybridization or amplification. Standard hybridization may be quantified by comparing the values obtained from normal subjects with values from an experiment in which a known amount of a substantially purified polynucleotide is used. Standard values obtained in this manner may be compared with values obtained from samples from patients who are symptomatic for a disorder. Deviation from standard values is used to establish the presence of a disorder.

[0173] Once the presence of a disorder is established and a treatment protocol is initiated, hybridization assays may be repeated on a regular basis to determine if the level of expression in the patient begins to approximate that which is observed in the normal subject. The results obtained from successive assays may be used to show the efficacy of treatment over a period ranging from several days to months.

[0174] With respect to cancer, the presence of an abnormal amount of transcript (either under- or overexpressed) in biopsied tissue from an individual may indicate a predisposition for the development of the disease, or may provide a means for detecting the disease prior to the appearance of actual clinical symptoms. A more definitive diagnosis of this type may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development or further progression of the cancer.

[0175] Additional diagnostic uses for oligonucleotides designed from the sequences encoding FLEXGEM may involve the use of PCR. These oligomers may be chemically synthesized, generated enzymatically, or produced in vitro. Oligomers will preferably contain a fragment of a polynucleotide encoding FLEXGEM, or a fragment of a polynucleotide complementary to the polynucleotide encoding FLEXGEM, and will be employed under optimized conditions for identification of a specific gene or condition. Oligomers may also be employed under less stringent conditions for detection or quantitation of closely related DNA or RNA sequences.

[0176] Methods which may also be used to quantify the expression of FLEXGEM include radiolabeling or biotinylating nucleotides, coamplification of a control nucleic acid, and interpolating results from standard curves. (See, e.g., Melby, P. C. et al. (1993) J. Immunol. Methods 159:235-244; Duplaa, C. et al. (1993) Anal. Biochem. 212:229-236.) The speed of quantification of multiple samples may be accelerated by running the assay in an ELISA format where the oligomer of interest is presented in various dilutions and a spectrophotometric or calorimetric response gives rapid quantification.

[0177] The polynucleotides are also useful for identifying individuals on the basis of minute biological samples, for example, by matching the restriction fragment length polymurphism (RFLP) pattern of a sample's DNA to that of an individual's DNA. The polynucleotides of the present invention can also be used to determine the actual base-by-base DNA sequence of selected portions of an individual's genome. These sequences can be used to prepare PCR primers for amplifying and isolating such selected DNA, which can then be sequenced. Using this technique, an individual can be identified through a unique set of DNA sequences. Once a unique ID database is established for an individual, positive identification of that individual can be made from extremely small tissue samples.

[0178] DNA-based identification techniques are critical in forensic technology. DNA sequences taken from very small biological samples such as tissues, e.g., hair or skin, or body fluids, e.g., blood, saliva, semen, etc., can be amplified using, e.g., PCR, to identify individuals. (See, e.g., Erlich, H., (1992) PCR Technology, Freeman and Co., New York, N.Y.). Similarly, polynucleotides of the present invention can be used as polymorphic markers.

[0179] There is also a need for reagents capable of identifying the source of a particular tissue.

[0180] Appropriate reagents can comprise, for example, DNA probes or primers prepared from the sequences of the present invention that are specific for particular tissues. Panels of such reagents can identify tissue by species and/or by organ type. In a similar fashion, these reagents can be used to screen tissue cultures for contamination.

[0181] The polynucleotides of the present invention can also be used as molecular weight markers on nucleic acid gels or Southern blots, as diagnostic probes for the presence of a specific mRNA in a particular cell type, in the creation of subtracted cDNA libraries which aid in the discovery of novel polynucleotides, in selection and synthesis of oligomers for attachment to an array or other support, and as an antigen to elicit an immune response.

[0182] In further embodiments, oligonucleotides or longer fragments derived from any of the polynucleotide sequences described herein may be used as targets in a microarray. The microarray can be used to monitor the expression level of large numbers of genes simultaneously and to identify genetic variants, mutations, and polymorphisms. This information may be used to determine gene function, to understand the genetic basis of a disorder, to diagnose a disorder, and to develop and monitor the activities of therapeutic agents.

[0183] Microarrays may be prepared, used, and analyzed using methods known in the art. (See, e.g., Brennan, T. M. et al. (1995) U.S. Pat. No. 5,474,796; Schena, M. et al. (1996) Proc. Natl. Acad. Sci. 93:10614-10619; Baldeschweiler et al. (1995) PCT application W0951251116; Shalon, D. et al. (1995) PCT application W095/35505; Heller, R. A. et al. (1997) Proc. Natl. Acad. Sci. 94:2150-2155; and Heller, M. J. et al. (1997) U.S. Pat. No. 5,605,662.)

[0184] For example, a chemical coupling procedure and an ink jet device can be used to synthesize array elements on the surface of a substrate. (See, e.g., Baldeschweiler, supra.) An array analogous to a dot or slot blot may also be used to arrange and link elements to the surface of a substrate using thermal, UV, chemical, or mechanical bonding procedures. A typical array may be produced by hand or using available methods and machines and contain any appropriate number of elements. After hybridization, nonhybridized probes are removed and a scanner used to determine the levels and patterns of fluorescence. The degree of complementarity and the relative abundance of each probe which hybridizes to an element on the microarray may be assessed through analysis of the scanned images.

[0185] Full-length cDNAs, Expressed Sequence Tags (ESTs), or fragments thereof may comprise the elements of the microarray. Fragments suitable for hybridization can be selected using software well known in the art such as LASERGENE software (DNASTAR). Full-length cDNAs, ESTs, or fragments thereof corresponding to one of the nucleotide sequences of the present invention, or selected at random from a cDNA library relevant to the present invention, are arranged on an appropriate substrate, e.g., a glass slide. The cDNA is fixed to the slide using, e.g., UV cross-linking followed by thermal and chemical treatments and subsequent drying. (See, e.g., Schena, M. et al. (1995) Science 270:467-470; Shalon, D. et al. (1996) Genome Res. 6:639-645.) Fluorescent probes are prepared and used for hybridization to the elements on the substrate. The substrate is analyzed by procedures described above.

Genetic Mapping

[0186] Gene identification and mapping are important in the investigation and treatment of almost all conditions, diseases, and disorders. Cancer, cardiovascular disease, Alzheimer's disease, arthritis, diabetes, and mental illnesses are of particular interest. Each of these conditions is more complex than the single gene defects of sickle cell anemia or cystic fibrosis, with select groups of genes being predictive of predisposition for a particular condition, disease, or disorder. For example, cardiovascular disease may result from malfunctioning receptor molecules that fail to clear cholesterol from the bloodstream, and diabetes may result when a particular individual's immune system is activated by an infection and attacks the insulin-producing cells of the pancreas. In some studies, Alzheimer's disease has been linked to a gene on chromosome 21; other studies predict a different gene and location. Mapping of disease genes is a complex and reiterative process and generally proceeds from genetic linkage analysis to physical mapping.

[0187] As a condition is noted among members of a family, a genetic linkage map traces parts of chromosomes that are inherited in the same pattern as the condition. Statistics link the inheritance of particular conditions to particular regions of chromosomes, as defined by RFLP or other markers. (See, for example, Lander, E. S. and Botstein, D. (1986) Proc. Natl. Acad. Sci. USA 83:7353-7357.) Occasionally, genetic markers and their locations are known from previous studies. More often, however, the markers are simply stretches of DNA that differ among individuals. Examples of genetic linkage maps can be found in various scientific journals or at the Online Mendelian Inheritance in Man (OMIM) World Wide Web site.

[0188] In another embodiment of the invention, FLEXGEM polynucleotide sequences may be used to generate hybridization probes useful in chromosomal mapping of naturally occurring genomic sequences. Either coding or noncoding polynucleotide sequences may be used, and in some instances, noncoding sequences may be preferable over coding sequences. For example, conservation of a coding sequence among members of a multi-gene family may potentially cause undesired cross hybridization during chromosomal mapping. The sequences may be mapped to a particular chromosome, to a specific region of a chromosome, or to artificial chromosome constructions, e.g., human artificial chromosomes (HACs), yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), bacterial P1 constructions, or single chromosome cDNA libraries. (See, e.g., Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355; Price, C. M. (1993) Blood Rev. 7:127-134; and Trask, B. J. (1991) Trends Genet. 7:149-15

[0189] Fluorescent in situ hybridization (FISH) may be correlated with other physical chromosome mapping techniques and genetic map data. (See, e.g., Meyers, supra, pp. 965-968.) Correlation between the location of a FLEXGEM polynucleotide sequence on a physical chromosomal map and a specific disorder, or a predisposition to a specific disorder, may help define the region of DNA associated with that disorder. The FLEXGEM polynucleotide sequences may also be used to detect polymorphisms that are genetically linked to the inheritance of a particular condition, disease, or disorder, or that modify susceptibility to a particular condition, disease, or disorder.

[0190] In situ hybridization of chromosomal preparations and genetic mapping techniques, such as linkage analysis using established chromosomal markers, may be used for extending existing genetic maps. Often the placement of a gene on the chromosome of another mammalian species, such as mouse, may reveal associated markers even if the number or arm of the corresponding human chromosome is not known. These new marker sequences can be mapped to human chromosomes and may provide valuable information to investigators searching for disease genes using positional cloning or other gene discovery techniques. Once a disease or syndrome has been crudely correlated by genetic linkage with a particular genomic region, e.g., ataxia-telangiectasia to 11q22-23, any sequences mapping to that area may represent associated or regulatory genes for further investigation. (See, e.g., Gatti, R. A. et al. (1988) Nature 336:577-580.) The nucleotide sequences of the subject invention may also be used to detect differences in chromosomal architecture due to translocation, inversion, etc., among normal, carrier, or affected individuals.

[0191] Once a disease-associated gene is mapped to a chromosomal region, the gene must be cloned in order to identify mutations or other alterations (e.g., translocations or inversions) that may be correlated with disease. This process requires a physical map of the chromosomal region containing the disease-gene of interest along with associated markers. A physical map is necessary for determining the nucleotide sequence and order of marker genes on a particular chromosomal region. Physical mapping techniques are well known in the art and require the generation of overlapping sets of cloned DNA fragments from a particular organelle, chromosome, or genome. These clones are analyzed to reconstruct and catalog their order. Once the position of a marker is determined, the DNA from that region is obtained by consulting the catalog and selecting clones from that region. The gene of interest is located through positional cloning techniques using hybridization or similar methods.

SNP Discovery and Detection

[0192] A further embodiment encompasses the use of polynucleotides encoding FLEXGEM toward the discovery of SNPs, preferably by microarray-based, high-throughput methods. Such methods would identify SNPs as variants, or alleles, of the polynucleotides of the invention among individuals or populations. Similar methods may then be used for genotyping and for the identification of those SNPs that are correlated with phenotypic characteristics such as disease risk or responsiveness to therapy. For example, the FLEXGEM alleles present in a disease-affected individual or population would be compared with those alleles observed in a control individual or population. The statistically significant occurrence of certain alleles in the diseased sample compared to the control sample indicates a correlation of these alleles with disease susceptibility or disease progression. Furthermore, the identification of SNPs present in the polynucleotides of the invention and the assignment of these SNPs to a particular chromosome map position would provide useful markers for human genetic studies and may provide a basis for haplotype analysis in human populations.

Screening

[0193] In another embodiment of the invention, FLEXGEM, its catalytic or immunogenic fragments, or oligopeptides thereof can be used for screening libraries of compounds in any of a variety of drug screening techniques. The fragment employed in such screening may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. The formation of binding complexes between FLEXGEM and the agent being tested may be measured.

[0194] Another technique for drug screening provides for high throughput screening of compounds having suitable binding affinity to the protein of interest. (See, e.g., Geysen, et al. (1984) PCT application WO84/03564.) In this method, large numbers of different small test compounds are synthesized on a solid substrate. The test compounds are reacted with FLEXGEM, or fragments thereof, and washed. Bound FLEXGEM is then detected by methods well known in the art. Purified FLEXGEM can also be coated directly onto plates for use in the aforementioned drug screening techniques. Alternatively, non-neutralizing antibodies can be used to capture the peptide and immobilize it on a solid support.

[0195] In another embodiment, one may use competitive drug screening assays in which neutralizing antibodies capable of binding FLEXGEM specifically compete with a test compound for binding FLEXGEM. In this manner, antibodies can be used to detect the presence of any peptide which shares one or more antigenic determinants with FLEXGEM.

[0196] In additional embodiments, the nucleotide sequences which encode FLEXGEM may be used in any molecular biology techniques that have yet to be developed, provided the new techniques rely on properties of nucleotide sequences that are currently known, including, but not limited to, such properties as the triplet genetic code and specific base pair interactions.

[0197] The examples below are provided to illustrate the subject invention and are not included for the purpose of limiting the invention.

EXAMPLES

[0198] I. Construction of cDNA Libraries

[0199] RNA was purchased from Clontech or isolated from tissues described in Table 4. Some tissues were homogenized and lysed in guanidinium isothiocyanate, while others were homogenized and lysed in phenol or in a suitable mixture of denaturants, such as TRIZOL (Life Technologies), a monophasic solution of phenol and guanidine isothiocyanate. The resulting lysates were centrifuged over CsCl cushions or extracted with chloroform. RNA was precipitated from the lysates with either isopropanol or sodium acetate and ethanol, or by other routine methods.

[0200] Phenol extraction and precipitation of RNA were repeated as necessary to increase RNA purity. In some cases, RNA was treated with DNase. For most libraries, poly(A+) RNA was isolated using oligo d(T)-coupled paramagnetic particles (Promega), OLIGOTEX latex particles (QIAGEN, Chatsworth Calif.), or an OLIGOTEX mRNA purification kit (QIAGEN). Alternatively, RNA was isolated directly from tissue lysates using other RNA isolation kits, e.g., the POLY(A)PURE mRNA purification kit (Ambion, Austin Tex.).

[0201] In some cases, Stratagene was provided with RNA and constructed the corresponding cDNA libraries. Otherwise, cDNA was synthesized and cDNA libraries were constructed with the UNIZAP vector system (Stratagene) or SUPERSCRIPT plasmid system (Life Technologies), using the recommended procedures or similar methods known in the art. (See, e.g., Ausubel, 1997, supra, units 5.1-6.6.) Reverse transcription was initiated using oligo d(T) or random primers. Synthetic oligonucleotide adapters were ligated to double stranded cDNA, and the cDNA was digested with the appropriate restriction enzyme or enzymes. For most libraries, the cDNA was size-selected (300-1000 bp) using SEPHACRYL S1000, SEPHAROSE CL2B, or SEPHAROSE CL4B column chromatography (Amersham Pharmacia Biotech) or preparative agarose gel electrophoresis. cDNAs were ligated into compatible restriction enzyme sites of the polylinker of a suitable plasmid, e.g., PBLUESCRIPT plasmid (Stratagene), pSPORT1 plasmid (Life Technologies), or pINCY (Incyte Pharmaceuticals, Palo Alto Calif.). Recombinant plasmids were transformed into competent E. coli cells including XL1-Blue, XL1-BlueMRF, or SOLR from Stratagene or DH5α, DH10B, or ElectroMAX DH10B from Life Technologies.

[0202] II. Isolation of cDNA Clones

[0203] Plasmids were recovered from host cells by in vivo excision using the UNIZAP vector system (Stratagene) or by cell lysis. Plasmids were purified using at least one of the following: a Magic or WIZARD Minipreps DNA purification system (Promega); an AGTC Miniprep purification kit (Edge Biosystems, Gaithersburg Md.); and QIAWELL 8 Plasmid, QIAWELL 8 Plus Plasmid, QIAWELL 8 Ultra Plasmid purification systems or the R.E.A.L. PREP 96 plasmid purification kit from QIAGEN. Following precipitation, plasmids were resuspended in 0.1 ml of distilled water and stored, with or without lyophilization, at 4° C.

[0204] Alternatively, plasmid DNA was amplified from host cell lysates using direct link PCR in a high-throughput format (Rao, V. B. (1994) Anal. Biochem. 216:1-14). Host cell lysis and thermal cycling steps were carried out in a single reaction mixture. Samples were processed and stored in 384-well plates, and the concentration of amplified plasmid DNA was quantified fluorometrically using PICOGREEN dye (Molecular Probes, Eugene Oreg.) and a FLUOROSKAN II fluorescence scanner (Labsystems Oy, Helsinki, Finland).

[0205] III. Sequencing and Analysis

[0206] cDNA sequencing reactions were processed using standard methods or high-throughput instrumentation such as the ABI CATALYST 800 (Perkin-Elmer) thermal cycler or the PTC-200 thermal cycler (MJ Research) in conjunction with the HYDRA microdispenser (Robbins Scientific) or the MICROLAB 2200 liquid transfer system (Hamilton). cDNA sequencing reactions were prepared using reagents provided by Amersham Pharmacia Biotech or supplied in ABI sequencing kits such as the ABI PRISM BIGDYE Terminator cycle sequencing ready reaction kit (Perkin-Elmer). Electrophoretic separation of cDNA sequencing reactions and detection of labeled polynucleotides were carried out using the MEGABACE 1000 DNA sequencing system (Molecular Dynamics); the ABI PRISM 373 or 377 sequencing system (Perkin-Elmer) in conjunction with standard ABI protocols and base calling software; or other sequence analysis systems known in the art. Reading frames within the cDNA sequences were identified using standard methods (reviewed in Ausubel, 1997, supra, unit 7.7). Some of the cDNA sequences were selected for extension using the techniques disclosed in Example V.

[0207] The polynucleotide sequences derived from cDNA sequencing were assembled and analyzed using a combination of software programs which utilize algorithms well known to those skilled in the art. Table 5 summarizes the tools, programs, and algorithms used and provides applicable descriptions, references, and threshold parameters. The first column of Table 5 shows the tools, programs, and algorithms used, the second column provides brief descriptions thereof, the third column presents appropriate references, all of which are incorporated by reference herein in their entirety, and the fourth column presents, where applicable, the scores, probability values, and other parameters used to evaluate the strength of a match between two sequences (the higher the score, the greater the homology between two sequences). Sequences were analyzed using MACDNASIS PRO software (Hitachi Software Engineering, South San Francisco Calif.) and LASERGENE software (DNASTAR). Polynucleotide and polypeptide sequence alignments were generated using the default parameters specified by the clustal algorithm as incorporated into the MEGALIGN multisequence alignment program (DNASTAR), which also calculates the percent identity between aligned sequences.

[0208] The polynucleotide sequences were validated by removing vector, linker, and polyA sequences and by masking ambiguous bases, using algorithms and programs based on BLAST, dynamic programming, and dinucleotide nearest neighbor analysis. The sequences were then queried against a selection of public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases, and BLOCKS using programs based on BLAST, FASTA, and BLIMPS. The sequences were assembled into full length polynucleotide sequences using programs based on Phred, Phrap, and Consed, and were screened for open reading frames using programs based on GeneMark, BLAST, and FASTA. The full length polynucleotide sequences were translated to derive the corresponding full length polypeptide sequences. The polypeptide sequences may be used to query databases such as the GenBank databases (described above), SwissProt, BLOCKS, PRINTS, Prosite, and Hidden Markov Model (HMM)-based protein family databases such as PFAM. HMM is a probabilistic approach which analyzes consensus primary structures of gene families. (See, e.g., Eddy, S. R. (1996) Curr. Opin. Str. Biol. 6:361-365.)

[0209] The programs described above for the assembly and analysis of full length polynucleotide and amino acid sequences were also used to identify polynucleotide sequence fragments from SEQ ID NO:21-40. Fragments from about 20 to about 4000 nucleotides which are useful in hybridization and amplification technologies were described in The Invention section above.

[0210] IV. Northern Analysis

[0211] Northern analysis is a laboratory technique used to detect the presence of a transcript of a gene and involves the hybridization of a labeled nucleotide sequence to a membrane on which RNAs from a particular cell type or tissue have been bound. (See, e.g., Sambrook, supra, ch. 7; Ausubel, 1995, supra, ch. 4 and 16.)

[0212] Analogous computer techniques applying BLAST were used to search for identical or related molecules in nucleotide databases such as GenBank or LIFESEQ (Incyte Pharmaceuticals). This analysis is much faster than multiple membrane-based hybridizations. In addition, the sensitivity of the computer search can be modified to determine whether any particular match is categorized as exact or similar. The basis of the search is the product score, which is defined as: $\frac{\% \quad {sequence}\quad {identity} \times \% \quad {maximum}\quad {BLAST}\quad {score}}{100}$

[0213] The product score takes into account both the degree of similarity between two sequences and the length of the sequence match. For example, with a product score of 40, the match will be exact within a 1% to 2% error, and, with a product score of 70, the match will be exact. Similar molecules are usually identified by selecting those which show product scores between 15 and 40, although lower scores may identify related molecules.

[0214] The results of northern analyses are reported as a percentage distribution of libraries in which the transcript encoding FLEXGEM occurred. Analysis involved the categorization of cDNA libraries by organ/tissue and disease. The organ/tissue categories included cardiovascular, dermatologic, developmental, endocrine, gastrointestinal, hematopoietic/immune, musculoskeletal, nervous, reproductive, and urologic. The disease/condition categories included cancer, inflammation/trauma, cell proliferation, neurological, and pooled. For each category, the number of libraries expressing the sequence of interest was counted and divided by the total number of libraries across all categories. Percentage values of tissue-specific and disease- or condition-specific expression are reported in Table 3.

[0215] V. Extension of FLEXGEM Encoding Polynucleotides

[0216] The full length nucleic acid sequences of SEQ ID NO:21-40 were produced by extension of an appropriate fragment of the full length molecule using oligonucleotide primers designed from this fragment. One primer was synthesized to initiate 5′ extension of the known fragment, and the other primer, to initiate 3′ extension of the known fragment. The initial primers were designed using OLIGO 4.06 software (National Biosciences), or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the target sequence at temperatures of about 68° C. to about 72° C. Any stretch of nucleotides which would result in hairpin structures and primer-primer dimerizations was avoided.

[0217] Selected human cDNA libraries were used to extend the sequence. If more than one extension was necessary or desired, additional or nested sets of primers were designed.

[0218] High fidelity amplification was obtained by PCR using methods well known in the art. PCR was performed in 96-well plates using the PTC-200 thermal cycler (MJ Research, Inc.). The reaction mix contained DNA template, 200 nmol of each primer, reaction buffer containing Mg²⁺, (NH₄)₂SO₄, and β-mercaptoethanol, Taq DNA polymerase (Amersham Pharmacia Biotech), ELONGASE enzyme (Life Technologies), and Pfu DNA polymerase (Stratagene), with the following parameters for primer pair PCI A and PCI B: Step 1: 94° C., 3 min; Step 2: 94° C., 15 sec; Step 3: 60° C., 1 min; Step 4: 68° C., 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times; Step 6: 68° C., 5 min; Step 7: storage at 4° C. In the alternative, the parameters for primer pair T7 and SK+ were as follows: Step 1: 94° C., 3 min; Step 2: 94° C., 15 sec 3: 57° C., 1 min; Step 4: 68° C., 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times; Step 6: 68° C., 5 min; Step 7: storage at 4° C.

[0219] The concentration of DNA in each well was determined by dispensing 100 μl PICOGREEN quantitation reagent (0.25% (v/v) PICOGREEN; Molecular Probes, Eugene Oreg.) dissolved in 1×TE and 0.5 μl of undiluted PCR product into each well of an opaque fluorimeter plate (Corning Costar, Acton Mass.), allowing the DNA to bind to the reagent. The plate was scanned in a Fluoroskan II (Labsystems Oy, Helsinki, Finland) to measure the fluorescence of the sample and to quantify the concentration of DNA. A 5 μl to 10 μl aliquot of the reaction mixture was analyzed by electrophoresis on a 1% agarose mini-gel to determine which reactions were successful in extending the sequence.

[0220] The extended nucleotides were desalted and concentrated, transferred to 384-well plates, digested with CviJI cholera virus endonuclease (Molecular Biology Research, Madison Wis.), and sonicated or sheared prior to religation into pUC 18 vector (Amersham Pharmacia Biotech). For shotgun sequencing, the digested nucleotides were separated on low concentration (0.6 to 0.8%) agarose gels, fragments were excised, and agar digested with Agar ACE (Promega). Extended clones were religated using T4 ligase (New England Biolabs, Beverly Mass.) into pUC 18 vector (Amersham Pharmacia Biotech), treated with Pfu DNA polymerase (Stratagene) to fill-in restriction site overhangs, and transfected into competent E. coli cells. Transformed cells were selected on antibiotic-containing media, individual colonies were picked and cultured overnight at 37° C. in 384-well plates in LB/2x carb liquid media.

[0221] The cells were lysed, and DNA was amplified by PCR using Taq DNA polymerase (Amersham Pharmacia Biotech) and Pfu DNA polymerase (Stratagene) with the following parameters: Step 1: 94° C., 3 min; Step 2: 94° C., 15 sec; Step 3: 60° C., 1 min; Step 4: 72° C., 2 min; Step 5: step 2, 3, and 4 repeated 29 times; Step 6: 72° C., 5 min; Step 7: storage at 4° C. DNA was quantified by PICOGREEN reagent (Molecular Probes) as described above. Samples with low DNA recoveries were reamplified using the same conditions as described above. Samples were diluted with 20% dimethysulphoxide (1:2, v/v), and sequenced using DYENAMIC energy transfer sequencing primers and the DYENAMIC DIRECT kit (Amersham Pharmacia Biotech) or the ABI PRISM BIGDYE Terminator cycle sequencing ready reaction kit (Perkin-Elmer).

[0222] In like manner, the nucleotide sequences of SEQ ID NO:21-40 are used to obtain 5′ regulatory sequences using the procedure above, oligonucleotides designed for such extension, and an appropriate genomic library.

[0223] VI. Labeling and Use of Individual Hybridization Probes

[0224] Hybridization probes derived from SEQ ID NO:21-40 are employed to screen cDNAs, genomic DNAs, or mRNAs. Although the labeling of oligonucleotides, consisting of about 20 base pairs, is specifically described, essentially the same procedure is used with larger nucleotide fragments. Oligonucleotides are designed using state-of-the-art software such as OLIGO 4.06 software (National Biosciences) and labeled by combining 50 pmol of each oligomer, 250 μCi of [γ-³²P] adenosine triphosphate (Amersham Pharmacia Biotech), and T4 polynucleotide kinase (DuPont NEN, Boston Mass.). The labeled oligonucleotides are substantially purified using a SEPHADEX G-25 superfine size exclusion dextran bead column (Amersham Pharmacia Biotech). An aliquot containing 10⁷ counts per minute of the labeled probe is used in a typical membrane-based hybridization analysis of human genomic DNA digested with one of the following endonucleases: Ase I, Bgl II, Eco RI, Pst I, Xbal, or Pvu II (DuPont NEN).

[0225] The DNA from each digest is fractionated on a 0.7% agarose gel and transferred to nylon membranes (Nytran Plus, Schleicher & Schuell, Durham N.H.). Hybridization is carried out for 16 hours at 40° C. To remove nonspecific signals, blots are sequentially washed at room temperature under increasingly stringent conditions up to 0.1 x saline sodium citrate and 0.5% sodium dodecyl sulfate. Hybridization patterns are visualized using autoradiography and compared.

[0226] VII. Transcript Image Analysis

[0227] Transcript images are generated as described in Seilhamer et al., “Comparative Gene Transcript Analysis,” U.S. Pat. No. 5,840,484, incorporated herein by reference.

[0228] VIII. Microarray Analysis

Probe Preparation from Tissue or Cell Samples

[0229] Total RNA is isolated from tissue samples using the guanidinium thiocyanate method and polyA⁺ RNA is purified using the oligo (dT) cellulose method. Each polyA⁺ RNA sample is reverse transcribed using MMLV reverse-transcriptase, 0.05 pg/μl oligo-dT primer (21 mer), 1X first strand buffer, 0.03 units/ul RNase inhibitor, 500 uM dATP, 500 uM dGTP, 500 uM dTTP, 40 uM dCTP, 40 uM dCTP-Cy3 (BDS) or dCTP-Cy5 (Amersham Pharmacia Biotech). The reverse transcription reaction is performed in a 25 ml volume containing 200 ng polyA⁺ RNA with GEMBRIGHT kits (Incyte). Specific control polyA⁺ RNAs are synthesized by in vitro transcription from non-coding yeast genomic DNA (W. Lei, unpublished). As quantitative controls, the control mRNAs at 0.002 ng, 0.02 ng, 0.2 ng, and 2 ng are diluted into reverse transcription reaction at ratios of 1:100,000, 1:10,000, 1:1000, 1:100 (w/w) to sample mRNA respectively. The control mRNAs are diluted into reverse transcription reaction at ratios of 1:3, 3:1, 1:10, 10:1, 1:25, 25:1 (w/w) to sample mRNA differential expression patterns. After incubation at 370 C for 2 hr, each reaction sample (one with Cy3 and another with Cy5 labeling) is treated with 2.5 ml of 0.5M sodium hydroxide and incubated for 20 minutes at 85° C. to the stop the reaction and degrade the RNA. Probes are purified using two successive CHROMA SPIN 30 gel filtration spin columns (CLONTECH Laboratories, Inc. (CLONTECH), Palo Alto Calif.) and after combining, both reaction samples are ethanol precipitated using 1 ml of glycogen (1 mg/ml), 60 ml sodium acetate, and 300 ml of 100% ethanol. The probe is then dried to completion using a SpeedVAC (Savant Instruments Inc., Holbrook N.Y.) and resuspended in 14 μl 5×SSC/0.2% SDS.

Microarray Preparation

[0230] Sequences of the present invention are used to generate array elements. Each array element is amplified from bacterial cells containing vectors with cloned cDNA inserts. PCR amplification uses primers complementary to the vector sequences flanking the cDNA insert. Array elements are amplified in thirty cycles of PCR from an initial quantity of 1-2 ng to a final quantity greater than 5 μg. Amplified array elements are then purified using SEPHACRYL-400 (Amersham Pharmacia Biotech).

[0231] Purified array elements are immobilized on polymer-coated glass slides. Glass microscope slides (Corning) are cleaned by ultrasound in 0. 1% SDS and acetone, with extensive distilled water washes between and after treatments. Glass slides are etched in 4% hydrofluoric acid (VWR Scientific Products Corporation (VWR), West Chester, Pa.), washed extensively in distilled water, and coated with 0.05% aminopropyl silane (Sigma) in 95% ethanol. Coated slides are cured in a 110° C. oven.

[0232] Array elements are applied to the coated glass substrate using a procedure described in U.S. Pat. No. 5,807,522, incorporated herein by reference. 1 μl of the array element DNA, at an average concentration of 100 ng/ul, is loaded into the open capillary printing element by a high-speed robotic apparatus. The apparatus then deposits about 5 nl of array element sample per slide.

[0233] Microarrays are UV-crosslinked using a STRATALINKER UV-crosslinker (Stratagene). Microarrays are washed at room temperature once in 0.2% SDS and three times in distilled water. Non-specific binding sites are blocked by incubation of microarrays in 0.2% casein in phosphate buffered saline (PBS) (Tropix, Inc., Bedford, Mass.) for 30 minutes at 60° C. followed by washes in 0.2% SDS and distilled water as before.

Hybridization

[0234] Hybridization reactions contain 9 μl of probe mixture consisting of 0.2 μg each of Cy3 and Cy5 labeled cDNA synthesis products in 5×SSC, 0.2% SDS hybridization buffer. The probe mixture is heated to 650 C for 5 minutes and is aliquoted onto the microarray surface and covered with an 1.8 cm² coverslip. The arrays are transferred to a waterproof chamber having a cavity just slightly larger than a microscope slide. The chamber is kept at 100% humidity internally by the addition of 140 μl of 5×SSC in a corner of the chamber. The chamber containing the arrays is incubated for about 6.5 hours at 60° C. The arrays are washed for 10 min at 45° C. in high stringency wash buffer (1×SSC, 0.1% SDS), three times for 10 minutes each at 45° C. in low stringency wash buffer (0. 1×SSC), and dried.

Detection

[0235] Reporter-labeled hybridization complexes are detected with a microscope equipped with an Innova 70 mixed gas 10 W laser (Coherent, Inc., Santa Clara Calif.) capable of generating spectral lines at 488 nm for excitation of Cy3 and at 632 nm for excitation of Cy5. The excitation laser light is focused on the array using a 20×microscope objective (Nikon, Inc., Melville N.Y.). The slide containing the array is placed on a computer-controlled X-Y stage on the microscope and raster-scanned past the objective. The 1.8 cm×1.8 cm array used in the present example is scanned with a resolution of 20 micrometers.

[0236] In two separate scans, a mixed gas multiline laser excites the two fluorophores sequentially. Emitted light is split, based on wavelength, into two photomultiplier tube detectors (PMT R1477, Hamamatsu Photonics Systems, Bridgewater N.J.) corresponding to the two fluorophores. Appropriate filters positioned between the array and the photomultiplier tubes are used to filter the signals. The emission maxima of the fluorophores used are 565 nm for Cy3 and 650 nm for Cy5. Each array is typically scanned twice, one scan per fluorophore using the appropriate filters at the laser source, although the apparatus is capable of recording the spectra from both fluorophores simultaneously.

[0237] The sensitivity of the scans is typically calibrated using the signal intensity generated by a cDNA control species added to the probe mix at a known concentration. A specific location on the array contains a complementary DNA sequence, allowing the intensity of the signal at that location to be correlated with a weight ratio of hybridizing species of 1:100,000. When two probes from different sources (e.g., representing test and control cells), each labeled with a different fluorophore, are hybridized to a single array for the purpose of identifying genes that are differentially expressed, the calibration is done by labeling samples of the calibrating cDNA with the two fluorophores and adding identical amounts of each to the hybridization mixture.

[0238] The output of the photomultiplier tube is digitized using a 12-bit RTI-835H analog-to-digital (A/D) conversion board (Analog Devices, Inc., Norwood, Mass.) installed in an IBM-compatible PC computer. The digitized data are displayed as an image where the signal intensity is mapped using a linear 20-color transformation to a pseudocolor scale ranging from blue (low signal) to red (high signal). The data is also analyzed quantitatively. Where two different fluorophores are excited and measured simultaneously, the data are first corrected for optical crosstalk (due to overlapping emission spectra) between the fluorophores using each fluorophore's emission spectrum.

[0239] A grid is superimposed over the fluorescence signal image such that the signal from each spot is centered in each element of the grid. The fluorescence signal within each element is then integrated to obtain a numerical value corresponding to the average intensity of the signal. The software used for signal analysis is the GEMTOOLS gene expression analysis program (Incyte).

[0240] IX. Complementary Polynucleotides

[0241] Sequences complementary to the FLEXGEM-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring FLEXGEM. Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the same procedure is used with smaller or with larger sequence fragments. Appropriate oligonucleotides are designed using OLIGO 4.06 software (National Biosciences) and the coding sequence of FLEXGEM. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5′ sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to the FLEXGEM-encoding transcript.

[0242] X. Expression of FLEXGEM

[0243] Expression and purification of FLEXGEM is achieved using bacterial or virus-based expression systems. For expression of FLEXGEM in bacteria, cDNA is subcloned into an appropriate vector containing an antibiotic resistance gene and an inducible promoter that directs high levels of cDNA transcription. Examples of such promoters include, but are not limited to, the trp-lac (tac) hybrid promoter and the T5 or T7 bacteriophage promoter in conjunction with the lac operator regulatory element. Recombinant vectors are transformed into suitable bacterial hosts, e.g., BL21(DE3). Antibiotic resistant bacteria express FLEXGEM upon induction with isopropyl beta-D-thiogalactopyranoside (IPTG). Expression of FLEXGEM in eukaryotic cells is achieved by infecting insect or mammalian cell lines with recombinant Autogrpahica californica nuclear polyhedrosis virus (AcMNPV), commonly known as baculovirus. The nonessential polyhedrin gene of baculovirus is replaced with cDNA encoding FLEXGEM by either homologous recombination or bacterial-mediated transposition involving transfer plasmid intermediates. Viral infectivity is maintained and the strong polyhedrin promoter drives high levels of cDNA transcription. Recombinant baculovirus is used to infect Spodoptera frugiperda (Sf9) insect cells in most cases, or human hepatocytes, in some cases. Infection of the latter requires additional genetic modifications to baculovirus. (See Engelhard, E. K. et al. (1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996) Hum. Gene Ther. 7:1937-1945.)

[0244] In most expression systems, FLEXGEM is synthesized as a fusion protein with, e.g., glutathione S-transferase (GST) or a peptide epitope tag, such as FLAG or 6-His, permitting rapid, single-step, affinity-based purification of recombinant fusion protein from crude cell lysates. GST, a 26-kilodalton enzyme from Schistosoma iaponicum, enables the purification of fusion proteins on immobilized glutathione under conditions that maintain protein activity and antigenicity (Amersham Pharmacia Biotech). Following purification, the GST moiety can be proteolytically cleaved from FLEXGEM at specifically engineered sites. FLAG, an 8-amino acid peptide, enables immunoaffinity purification using commercially available monoclonal and polyclonal anti-FLAG antibodies (Eastman Kodak). 6-His, a stretch of six consecutive histidine residues, enables purification on metal-chelate resins (QIAGEN). Methods for protein expression and purification are discussed in Ausubel (1995, supra, ch 10 and 16). Purified FLEXGEM obtained by these methods can be used directly in the following activity assay.

[0245] XI. Functional Assays

[0246] FLEXGEM function is assessed by expressing the sequences encoding FLEXGEM at physiologically elevated levels in mammalian cell culture systems. cDNA is subcloned into a mammalian expression vector containing a strong promoter that drives high levels of cDNA expression. Vectors of choice include pCMV SPORT (Life Technologies) and pCR3.1 (Invitrogen, Carlsbad Calif.), both of which contain the cytomegalovirus promoter. 5-10 μg of recombinant vector are transiently transfected into a human cell line, preferably of endothelial or hematopoietic origin, using either liposome formulations or electroporation. 1-2 μg of an additional plasmid containing sequences encoding a marker protein are co-transfected. Expression of a marker protein provides a means to distinguish transfected cells from nontransfected cells and is a reliable predictor of cDNA expression from the recombinant vector. Marker proteins of choice include, e.g., Green Fluorescent Protein (GFP; Clontech), CD64, or a CD64-GFP fusion protein. Flow cytometry (FCM), an automated, laser optics-based technique, is used to identify transfected cells expressing GFP or CD64-GFP and to evaluate the apoptotic state of the cells and other cellular properties. FCM detects and quantifies the uptake of fluorescent molecules that diagnose events preceding or coincident with cell death. These events include changes in nuclear DNA content as measured by staining of DNA with propidium iodide; changes in cell size and granularity as measured by forward light scatter and 90 degree side light scatter; down-regulation of DNA synthesis as measured by decrease in bromodeoxyuridine uptake; alterations in expression of cell surface and intracellular proteins as measured by reactivity with specific antibodies; and alterations in plasma membrane composition as measured by the binding of fluorescein-conjugated Annexin V protein to the cell surface. Methods in flow cytometry are discussed in Ormerod, M. G. (1994) Flow Cytometry, Oxford, New York N.Y.

[0247] The influence of FLEXGEM on gene expression can be assessed using highly purified populations of cells transfected with sequences encoding FLEXGEM and either CD64 or CD64-GFP. CD64 and CD64-GFP are expressed on the surface of transfected cells and bind to conserved regions of human immunoglobulin G (IgG). Transfected cells are efficiently separated from nontransfected cells using magnetic beads coated with either human IgG or antibody against CD64 (DYNAL, Lake Success N.Y.). mRNA can be purified from the cells using methods well known by those of skill in the art. Expression of mRNA encoding FLEXGEM and other genes of interest can be analyzed by northern analysis or microarray techniques.

[0248] XII. Production of FLEXGEM Specific Antibodies

[0249] FLEXGEM substantially purified using polyacrylamide gel electrophoresis (PAGE; see, e.g., Harrington, M. G. (1990) Methods Enzymol. 182:488-495), or other purification techniques, is used to immunize rabbits and to produce antibodies using standard protocols.

[0250] Alternatively, the FLEXGEM amino acid sequence is analyzed using LASERGENE software (DNASTAR) to determine regions of high immunogenicity, and a corresponding oligopeptide is synthesized and used to raise antibodies by means known to those of skill in the art. Methods for selection of appropriate epitopes, such as those near the C-terminus or in hydrophilic regions are well described in the art. (See, e.g., Ausubel, 1995, supra, ch. 11.)

[0251] Typically, oligopeptides 15 residues in length are synthesized using an ABI 431A peptide synthesizer (Perkin-Elmer) using fmoc-chemistry and coupled to KLH (Sigma-Aldrich, St. Louis Mo.) by reaction with N-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) to increase immunogenicity. (See, e.g., Ausubel, 1995, supra.) Rabbits are immunized with the oligopeptide-KLH complex in complete Freund's adjuvant. Resulting antisera are tested for antipeptide activity by, for example, binding the peptide to plastic, blocking with 1% BSA, reacting with rabbit antisera, washing, and reacting with radio-iodinated goat anti-rabbit IgG.

[0252] XIII. Purification of Naturally Occurring FLEXGEM Using Specific Antibodies

[0253] Naturally occurring or recombinant FLEXGEM is substantially purified by immunoaffinity chromatography using antibodies specific for FLEXGEM. An immunoaffinity column is constructed by covalently coupling anti-FLEXGEM antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.

[0254] Media containing FLEXGEM are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of FLEXGEM (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/FLEXGEM binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and FLEXGEM is collected.

[0255] XIV. Identification of Molecules Which Interact with FLEXGEM

[0256] FLEXGEM, or biologically active fragments thereof, are labeled with 125I Bolton-Hunter reagent. (See, e.g., Bolton et al. (1973) Biochem. J. 133:529.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled FLEXGEM, washed, and any wells with labeled FLEXGEM complex are assayed. Data obtained using different concentrations of FLEXGEM are used to calculate values for the number, affinity, and association of FLEXGEM with the candidate molecules.

[0257] Alternatively, molecules interacting with FLEXGEM are analyzed using the yeast two-hybrid system as described in Fields, S. and Song, O. (1989) Nature 340:245-246, or using commercially available kits based on the two-hybrid system, such as the MATCHMAKER system (CLONTECH).

[0258] Various modifications and variations of the described methods and systems of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims. TABLE I1 Protein SEQ Nucleotide Clone ID NO: SEQ ID NO: ID Library Fragments 1 21 1841446 COLNNOT07 869045R6 (LUNGAST01), 1251087F6 (LUNGFET03), 1574990F6 (LNODNOT03), 1841446H1 (COLNNOT07), 1860233F6 and 1860233T6 (PROSNOT18), 4999024H1 (MYEPTXT02) 2 22 1850310 LUNGFET03 045378H1 (CORNNOT01), 178670H1 (PLACNOB01), 1731750F6 (BRSTTUT08), 1850310F6 (LUNGFET03), 1850310H1, 1850310T6, and 1850465F6 (LUNGFET03), 2171482F6 (ENDCNOT03), 2771742H1 (COLANOT02), 3689884H1 (HEAANOT01), 5388293H1 (BRAINOT19) 3 23 1887020 BLADTUT07 997447R1 (KIDNTUT01), 1603324F6 (LUNGNOT15), 1887020F6 and 1887020H1 (BLADTUT07), 3149213H1 (ADRENON04), 3415447H1 (PTHYNOT04), 3472602H1 (LUNGNOT27), 5043722H2 (PLACFER01), 5300608H1 (MUSCNOT11) 4 24 1911421 CONNTUT01 276895H1 and 278868H1 (TESTNOT03), 1911421H1 and 1911421T6 (CONNTUT01), 2458650F6 (ENDANOT01) 5 25 1911910 CONNTUT01 1321511F1 (BLADNOT04), 1461469R1 (PANCNOT04), 1842277T6 (COLNNOT07), 1911910F6 and 1911910H1 (CONNTUT01) 6 26 1928920 BRSTNOT02 1928920H1 (BRSTNOT02), 637943F1 (BRSTNOT03), 1260702R1 (SYNORAT05), 1461588T1 (PANCNOT04), 1754346F6 (LIVRTUT01), 2906971F6 (THYMNOT05), 3368705F6 (CONNTUT04) 7 27 2170846 ENDCNOT03 2170846F6 and 2170846H1 (ENDCNOT03), 2828087T6 (TLYMNOT03), SBLA00670F1 8 28 2176361 ENDCNOT03 660127X300D2 (BRAINOT03), 1212293R6 (BRSTTUT01), 1310486T1 (COLNFET02), 1841413H1 (COLNNOT07), 2176361H1 and 2176361T6 (ENDCNOT03), 3212993T6 (BLADNOT08) 9 29 2212732 SINTFET03 285007F1 (EOSIHET02), 1002395R1 (BRSTNOT03), 2212732H1 (SINTFET03), 2361923R6 (LUNGFET05), 2708785T6 (PONSAZT01) , 3605611H1 (LUNGNOT30) 10 30 2303457 BRSTNOT05 1573212F6 (LNODNOT03), 2303457H1 (BRSTNOT05), 2539058F6 (BONRTUT01), 3605342H1 (LUNGNOT30) 11 31 2317552 OVARNOT02 409183F1 (EOSIHET02), 863694R1 and 863694T1 (BRAITUT03), 913694H1 (STOMNOT02), 1319506T1 (BLADNOT04), 2317552H1 (OVARNOT02), 2604101H1 (LUNGTUT07) 12 32 2416366 HNT3AZT01 228599F1 (PANCNOT01), 1384425T1 (BRAITUT08), 1636237F6 (UTRSNOT06), 2416366H1 (HNT3AZT01), 2600055F6 (UTRSNOT10), 2868081F6 and 2868081T6 (KIDNNOT20) 13 33 2472980 THP1NOT03 034079R6 and 034079T6 (THP1NOB01), 1595315F6 (BRAINOT14), 2472980H1 (THP1NOT03), 2596942T6 (OVARTUT02), 3852779T6 (BRAITUT12), 4754861H1 (BRAHNOT01) 14 34 2541640 BONRTUT01 621985F1 and 621985R6 (PGANNOT01), 2541640F6 and 2541640H1 (BONRTUT01), 5314848H1 (KIDETXS02) 15 35 2695204 UTRSNOT12 927986R1 (BRAINOT04), 1854107F6 (HNT3AZT01), 2695204H1 (UTRSNOT12), 3464157F6 (293TF2T01), 3596821H1 (FIBPNOT01) 16 36 2805526 BLADTUT08 161563F1 (ADENINB01), 1275596F6 (TESTTUT02), 1508327F6 (LUNGNOT14), 1965858H1 (BRSTNOT04), 2207901F6 (SINTFET03), 2805526H1 (BLADTUT08), 3602953F6 (DRGTNOT01) 17 37 2850382 BRSTTUT13 1217874T1 (NEUTGMT01), 1267183F1 (BRAINOT09), 1515874F1 (PANCTUT01), 2234171F6 (PANCTUT02), 2850382H1 (BRSTTUT13), 3085014H1 (HEAONOT03) 18 38 2929276 TLYMNOT04 2929276H1 (TLYMNOT04) 136458R1 (SYNORAB01), 149704R1 (FIBRNGT02), 1231828X27 (BRAITUT01), 1236144F1 (LUNGFET03), 1351889F1 (LATRTUT02), 1381121F1 (BRAITUT08), 2509933F6 (CONUTUT01), 2819412H1 (BRSTNOT14), 3029720H1 (HEARFET02), 4897855H1 (OVARDIT01) 19 39 3033039 TLYMNOT05 3033039H1 (TLYMNOT05), 1235556F1 and 1251391F1 (LUNGFET03), 1261615H1 (SYNORAT05), 1413023F6 (BRAINOT12), 1425681T1 (BEPINON01), 1498538F1 (SINTBST01), 1562136F1 (SPLNNOT04), 1901480T6 (BLADTUT06), 2170485F6 (ENDCNOT03), 2454842F6 (ENDANOT01), 2915250H1 (THYMFET03), 4248873H1 (BRADDIR01), 4721266H1 (BRAIHCT02), 4939383H1 (BRAIFEN03), 5094946H1 (EPIMNON05), 5151444H1 (HEARFET03) 20 40 3039890 BRSTNOT16 1535201T1 (SPLNNOT04), 1576892T1 (LNODNOT03), 2517426H1 (BRAITUT21), 2715705H1 (THYRNOT09), 3039890H1 and 3039890T6 (BRSTNOT16), SBMA03391F1, SBMA02852F1, SBMA02370F1

[0259] TABLE 2 Amino Potential Polypeptide Acid Glycosylation Seq ID NO: Residues Potential Phosphorylation Sites Sites 1 349 S53 T114 T147 S11 S15 S141 S149 N250 T156 S225 T252 S299 T313 T342 S343 2 169 T39 T88 T71 S119 T128 N126 3 316 T85 S194 T196 S80 T230 4 220 T39 S10 T169 S77 S205 S208 N15 5 235 S13 T153 S128 S182 S134 T187 Y155 6 487 S235 S3 S183 T149 L448 7 212 S59 T84 Y147 N57 N206 8 241 T167 S168 S48 T79 S129 S7 T36 S66 T86 T219 9 375 T67 T111 S123 S132 T317 S8 S81 T173 S355 Y19 10 429 S257 S40 T90 S117 S232 S234 S236 T242 N88 N173 N283 T252 S256 S387 11 329 S8 T10 S29 S47 S49 S57 S227 S260 S314 N64 N114 N122 N250 T86 S124 S277 12 476 S195 T246 T453 S459 S10 S20 S22 S34 N3 N376 S55 S57 S72 S77 S95 S97 S118 S122 S137 S168 S184 S211 T263 S330 T357 T392 S415 S443 S64 S83 S86 S106 S197 T308 T382 S431 T437 S438 13 366 S330 T119 T188 S60 T65 S140 S185 N39 N161 N202 N269 T298 S305 T326 T337 S344 S357 N273 N348 N352 14 152 T81 T148 S45 N28 15 233 T87 S18 T172 S186 S189 S203 N130 16 357 T118 S189 T62 T80 S115 T126 S247 S273 S328 S337 17 251 T48 T224 S106 T114 S126 S161 T180 T202 S235 18 105 S49 T69 T98 S15 S34 19 876 S119 S179 T180 S226 T262 T386 S450 N397 N644 S494 S529 T542 S560 S585 T689 S716 T728 S770 S804 T35 T110 T123 S238 T248 T457 S574 S634 S693 T702 S733 S745 S806 20 505 S377 S4 T129 S179 T338 T339 T413 N320 N477 T484 S179 S208 S377 S439 S463 S474 S491

[0260] TABLE 3 Nucleotide Useful Tissue Expression Disease or Condition SEQ ID NO: Fragment (Fraction of Total) (Fraction of Total) Vector 21 596-640 Reproductive (0.235) Cancer (0.432) pSPORT1 Hematopoietic/Immune (0.148) Inflammation (0.284) Cardiovascular (0.136) Cell Proliferation (0.222) 22 1380-1424 Reproductive (0.296) Cancer (0.426) pINCY Nervous (0.167) Inflammation (0.222) Gastrointestinal (0.130) Cell Proliferation (0.204) 23 273-317 Reproductive (0.200) Cancer (0.475) Cell pINCY Cardiovascular (0.175) Proliferation (0.175) Nervous (0.175) Inflammation (0.175) 24 111-155 Reproductive (0.215) Cancer (0.430) pINCY Cardiovascular (0.139) Inflammation (0.241) Nervous (0.139) Cell Proliferation (0.228) 25 217-261 Nervous (0.351) Cancer (0.405) Cell pINCY Gastrointestinal (0.135) Proliferation (0.216) Reproductive (0.135) Inflammation (0.189) 26 543-587 Reproductive (0.237) Cancer (0.475) pSPORT1 Gastrointestinal (0.175) Inflammation (0.237) Nervous (0.150) Cell Proliferation (0.225) 27 138-182 Reproductive (0.250) Cancer (0.500) Cell pINCY Cardiovascular (0.200) Proliferation (0.250) Gastrointestinal (0.150) 28 1029-1073 Reproductive (0.325) Cancer (0.450) Cell pINCY Nervous (0.125) Proliferation (0.200) Urologic (0.125) Inflammation (0.150) 29 434-478 Nervous (0.250) Cancer (0.444) pINCY Reproductive (0.222) Inflammation (0.306) Gastrointestinal (0.139) Cell Proliferation (0.194) 30 327-372 Reproductive (0.333) Cancer (0.389) pSPORT1 Nervous (0.278) Inflammation (0.278) Cardiovascular (0.111) Cell Proliferation (0.111) 31  992-1036 Nervous (0.235) Cancer (0.382) pSPORT1 Reproductive (0.235) Inflammation (0.255) Gastrointestinal (0.157) Cell Proliferation (0.235) 32 325-369 Nervous (0.250) Cancer (0.484) pINCY Reproductive (0.219) Inflammation (0.203) Hematopoietic/Immune (0.172) Cell Proliferation (0.141) 33 336-380 Reproductive (0.219) Cancer (0.438) pINCY Developmental (0.156) Inflammation (0.250) Hematopoietic/Immune (0.156) Cell Proliferation (0.219) 34 109-153 Reproductive (0.364) Cancer (0.818) pINCY Gastrointestinal (0.273) Cell Proliferation (0.091) Musculoskeletal (0.182) Trauma (0.091) 35 597-641 Nervous (0.211) Cancer (0.395) pINCY Reproductive (0.211) Inflammation (0.263) Hematopoietic/Immune (0.132) Cell Proliferation (0.132) 36 921-965 Reproductive (0.324) Cancer (0.514) pINCY Nervous (0.135) Inflammation (0.162) Developmental (0.108) Cell Proliferation (0.135) 37 381-425 Reproductive (0.269) Cancer (0.436) pINCY Nervous (0.192) Inflammation (0.192) Hematopoietic/Immune (0.141) Cell Proliferation (0.167) 38 109-153 Reproductive (0.281) Cancer (0.412) pINCY Hematopoietic/Immune (0.132) Inflammation (0.333) Cardiovascular (0.114) Cell Proliferation (0.158) 39 433-477 Nervous (0.206) Cancer (0.477) Cell pINCY Reproductive (0.206) Proliferation (0.224) Cardiovascular (0.150) Inflammation (0.206) 40 596-640 Reproductive (0.239) Cancer (0.439) pINCY Nervous (0.181) Inflammation (0.219) Hematopoietic/Immune (0.129) Cell Proliferation (0.194)

[0261] TABLE 4 Nucleotide SEQ ID NO: Library Library Comment 21 COLNNOT07 Library was constructed using RNA isolated from colon tissue removed from a 60-year-old Caucasian male during a left hemicolectomy. 22 LUNGFET03 Library was constructed using RNA isolated from lung tissue removed from a Caucasian female fetus, who died at 20 weeks' gestation. 23 BLADTUT07 Library was constructed using RNA isolated from bladder tumor tissue removed from the anterior bladder wall of a 58-year-old Caucasian male during a radical cystectomy, radical prostatectomy, and gastrostomy. Pathology indicated a grade 3 transitional cell carcinoma in the left lateral bladder. Patient history included angina and emphysema. Family history included acute myocardial infarction, atherosclerotic coronary artery disease, and type II diabetes. 24 CONNTUT01 Library was constructed using RNA isolated from a soft tissue tumor removed from the clival area of the skull of a 30-year-old Caucasian female. Pathology indicated chondroid chordoma with neoplastic cells reactive for keratin. 25 CONNTUT01 Library was constructed using RNA isolated from a soft tissue tumor removed from the clival area of the skull of a 30-year-old Caucasian female. Pathology indicated chondroid chordoma with neoplastic cells reactive for keratin. 26 BRSTNOT02 Library was constructed using RNA isolated from diseased breast tissue removed from a 55-year-old Caucasian female during a unilateral extended simple mastectomy. Pathology indicated proliferative fibrocysytic changes characterized by apocrine metaplasia, sclerosing adenosis, cyst formation, and ductal hyperplasia without atypia. Pathology for the associated tumor tissue indicated an invasive grade 4 mammary adenocarcinoma. Patient history included atrial tachycardia and a benign neoplasm. Family history included cardiovascular and cerebrovascular disease. 27 ENDCNOT03 Library was constructed using RNA isolated from dermal microvascular endothelial cells removed from a neonatal Caucasian male. 28 ENDCNOT03 Library was constructed using RNA isolated from dermal microvascular endothelial cells removed from a neonatal Caucasian male. 29 SINTFET03 Library was constructed using RNA isolated from small intestine tissue removed from a Caucasian female fetus, who died at 20 weeks' gestation. 30 BRSTNOT05 Library was constructed using RNA isolated from breast tissue removed from a 58-year-old Caucasian female during a unilateral extended simple mastectomy. Pathology for the associated tumor tissue indicated multicentric invasive grade 4 lobular carcinoma. Patient history included skin cancer, rheumatic heart disease, osteoarthritis, and tuberculosis. Family history included cerebrovascular and cardiovascular disease, breast and prostate cancer, and type I diabetes. 31 OVARNOT02 Library was constructed using RNA isolated from ovarian tissue removed from a 59-year-old Caucasian female who died of a myocardial infarction. Patient history included cardiomyopathy, coronary artery disease, previous myocardial infarctions, hypercholesterolemia, hypotension, and arthritis. 32 HNT3AZT01 Library was constructed using RNA isolated from the hNT2 cell line (derived from a human teratocarcinoma that exhibited properties characteristic of a committed neuronal precursor). Cells were treated for three days with 0.35 micromolar 5-aza-2′-deoxycytidine (AZ). 33 THP1NOT03 Library was constructed using polyA RNA isolated from untreated THP-1 cells. THP-1 (ATCC TIB 202) is a human promonocyte line derived from the peripheral blood of a 1-year-old Caucasian male with acute monocytic leukemia (ref: Int. J. Cancer (1980) 26:171). 34 BONRTUT01 Library was constructed using RNA isolated from rib tumor tissue removed from a 16-year-old Caucasian male during a rib osteotomy and a wedge resection of the lung. Pathology indicated a metastatic grade 3 (of 4) osteosarcoma, forming a mass involving the chest wall. 35 UTRSNOT12 Library was constructed using RNA isolated from uterine myometrial tissue removed from a 41-year-old Caucasian female during a vaginal hysterectomy with dilation and curettage. The endometrium was secretory and contained fragments of endometrial polyps. Benign endo- and ectocervical mucosa were identified in the endocervix. Pathology for the associated tumor tissue indicated uterine leiomyoma. 36 BLADTUT08 Library was constructed using RNA isolated from bladder tumor tissue removed from a 72-year-old Caucasian male during a radical cystectomy and prostatectomy. Pathology indicated an invasive grade 3 (of 3) transitional cell carcinoma in the right bladder base. Family history included myocardial infarction, cerebrovascular disease, and brain cancer. 37 BRSTTUT13 Library was constructed using RNA isolated from breast tumor tissue removed from the right breast of a 46-year-old Caucasian female during a unilateral extended simple mastectomy with breast reconstruction. Pathology indicated an invasive grade 3 adenocarcinoma, ductal type with apocrine features and greater than 50% intraductal component. Patient history included breast cancer. 38 TLYMNOT04 Library was constructed using 0.5 micrograms of polyA RNA isolated from activated Th1 cells. These cells were differentiated from umbilical cord CD4 T cells with IL-12 and B7-transfected COS cells, and then activated for six hours with anti-CD3 and anti-CD28 antibodies. 39 TLYMNOT05 Library was constructed using polyA RNA isolated from nonactivated Th2 cells. These cells were differentiated from umbilical cord CD4 T cells with IL-4 in the presence of anti-IL-12 antibodies and B7-transfected COS cells. 40 BRSTNOT16 Library was constructed using RNA isolated from diseased breast tissue removed from the right breast of a 59-year-old Caucasian female during a unilateral extended simple mastectomy. Pathology indicated papillomatosis. Pathology for the associated tumor tissue indicated an invasive lobular carcinoma with extension into ducts, forming an ill-defined mass situated in the biopsy cavity site. Multiple axillary lymph nodes were negative for tumor. Prior right breast biopsy indicated invasive grade 3, nuclear grade 3, invasive and in situ ductal carcinoma. Estrogen and progesterone immunostains were positive in the neoplastic cells. Patient history included liver cirrhosis, esophageal ulcer, hyperlipidemia, and neuropathy.

[0262] TABLE 5 Program Description Reference Parameter Threshold ABI FACTURA A program that removes vector Perkin-Elmer Applied Biosystems, sequences and masks ambiguous bases Foster City, CA. in nucleic acid sequences. ABI/PARACEL FDF A Fast Data Finder useful in Perkin-Elmer Applied Biosystems, Mismatch <50% comparing and annotating amino acid Foster City, CA; Paracel Inc., Pasadena, CA. or nucleic acid sequences. ABI AutoAssembler A program that assembles nucleic Perkin-Elmer Applied Biosystems, acid sequences. Foster City, CA. BLAST A Basic Local Alignment Search Tool Altschul, S. F. et al. (1990) J. Mol. Biol. ESTs: Probability value = 1.0E−8 useful in sequence similarity search for 215:403-410; Altschul, S.F. et al. (1997) or less amino acid and nucleic acid sequences. Nucleic Acids Res. 25: 3389-3402. Full Length sequences: BLAST includes five functions: blastp, Probability value = 1.0E−10 or less blastn, blastx, tblastn, and tblastx. FASTA A Pearson and Lipman algorithm that Pearson, W. R. and D. J. Lipman (1988) Proc. ESTs: fasta E value = 1.06E−6 searches for similarity between a query Natl. Acad Sci. 85:2444-2448; Pearson, W. R. Assembled ESTs: fasta Identity = sequence and a group of sequences of (1990) Methods Enzymol. 183: 63-98; and 95% or greater and Match the same type. FASTA comprises as Smith, T. F. and M. S. Waterman (1981) Adv. length = 200 bases or greater; fastx least five functions: fasta, tfasta, fastx, Appl. Math. 2:482-489. E value = 1.0E−8 or less tfastx, and ssearch. Full Length sequences: fastx score = 100 or greater BLIMPS A BLocks IMProved Searcher that Henikoff, S and J. G. Henikoff, Nucl. Acid Res., Score = 1000 or greater; Ratio of matches a sequence against those in 19:6565-72, 1991. J. G. Henikoff and S. Score/Strength = 0.75 or larger; BLOCKS and PRINTS databases to Henikoff (1996) Methods Enzymol. 266:88-105; and Probability value = 1.0E−3 or search for gene families, sequence and Attwood, T. K. et al. (1997) J. Chem. Inf. less homology, and structural fingerprint Comput. Sci. 37: 417-424. PFAM A Hidden Markov Models-based Krogh, A. et al. (1994) J. Mol. Biol., 235:1501- Score = 10-50 bits, depending on application useful for protein family 1531; Sonnhammer, E. L. L. et al. (1988) individual protein families search. Nucleic Acids Res. 26:320-322. ProfileScan An algorithm that searches for structural Gribskov, M. et al. (1988) CABIOS 4:61-66; Score = 4.0 or greater and sequence motifs in protein Gribskov, et al. (1989) Methods Enzymol. sequences that match sequence patterns 183:146-159; Bairoch, A. et al. (1997) Nucleic defined in Prosite. Acids Res. 25: 217-221. Phred A base-calling algorithm that examines Ewing, B. et al. (1998) Genome automated sequencer traces with high Res. 8:175-185; Ewing, B. and P. sensitivity and probability. Green (1998) Genome Res. 8:186- 194. Phrap A Phils Revised Assembly Program Smith, T. F. and M. S. Waterman (1981) Adv. Score = 120 or greater; Match including SWAT and CrossMatch, Appl. Math. 2:482-489; Smith, T. F. and M. S. length = 56 or greater programs based on efficient Waterman (1981) J. Mol. Biol. 147:195-197; implementation of the Smith-Waterman and Green, P., University of Washington, algorithm, useful in searching sequence Seattle, WA. homology and assembling DNA sequences. Consed A graphical tool for viewing and editing Gordon, D. et al. (1998) Genome Phrap assemblies Res. 8:195-202. SPScan A weight matrix analysis program that Nielson, H. et al. (1997) Protein Engineering Score = 5 or greater scans protein sequences for the presence 10:1-6; Claverie, J. M. and S. Audic (1997) of secretory signal peptides. CABIOS 12: 431-439. Motifs A program that searches amino acid Bairoch et al. supra; Wisconsin sequences for patterns that matched Package Program Manual, version those defined in Prosite. 9, page M51-59, Genetics Computer Group, Madison, WI.

[0263]

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 40 <210> SEQ ID NO 1 <211> LENGTH: 349 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1841446 <400> SEQUENCE: 1 Met Ala Lys Ala Gly Asp Lys Ser Ser Ser Ser Gly Lys Lys Ser 1 5 10 15 Leu Lys Arg Lys Ala Ala Ala Glu Glu Leu Gln Glu Ala Ala Gly 20 25 30 Ala Gly Asp Gly Ala Thr Glu Asn Gly Val Gln Pro Pro Lys Ala 35 40 45 Ala Ala Phe Pro Pro Gly Phe Ser Ile Ser Glu Ile Lys Asn Lys 50 55 60 Gln Arg Arg His Leu Met Phe Thr Arg Trp Lys Gln Gln Gln Arg 65 70 75 Lys Glu Lys Leu Ala Ala Lys Lys Lys Leu Lys Lys Glu Arg Glu 80 85 90 Ala Leu Gly Asp Lys Ala Pro Pro Lys Pro Val Pro Lys Thr Ile 95 100 105 Asp Asn Gln Arg Val Tyr Asp Glu Thr Thr Val Asp Pro Asn Asp 110 115 120 Glu Glu Val Ala Tyr Asp Glu Ala Thr Asp Glu Phe Ala Ser Tyr 125 130 135 Phe Asn Lys Gln Thr Ser Pro Lys Ile Leu Ile Thr Thr Ser Asp 140 145 150 Arg Pro His Gly Arg Thr Val Arg Leu Cys Glu Gln Leu Ser Thr 155 160 165 Val Ile Pro Asn Ser His Val Tyr Tyr Arg Arg Gly Leu Ala Leu 170 175 180 Lys Lys Ile Ile Pro Gln Cys Ile Ala Arg Asp Phe Thr Asp Leu 185 190 195 Ile Val Ile Asn Glu Asp Arg Lys Thr Pro Asn Gly Leu Ile Leu 200 205 210 Ser His Leu Pro Asn Gly Pro Thr Ala His Phe Lys Met Ser Ser 215 220 225 Val Arg Leu Arg Lys Glu Ile Lys Arg Arg Gly Lys Asp Pro Thr 230 235 240 Glu His Ile Pro Glu Ile Ile Leu Asn Asn Phe Thr Thr Arg Leu 245 250 255 Gly His Ser Ile Gly Arg Met Phe Ala Ser Leu Phe Pro His Asn 260 265 270 Pro Gln Phe Ile Gly Arg Gln Val Ala Thr Phe His Asn Gln Arg 275 280 285 Asp Tyr Ile Phe Phe Arg Phe His Arg Tyr Ile Phe Arg Ser Glu 290 295 300 Lys Lys Val Gly Ile Gln Glu Leu Gly Pro Arg Phe Thr Leu Lys 305 310 315 Leu Arg Ser Leu Gln Lys Gly Thr Phe Asp Ser Lys Tyr Gly Glu 320 325 330 Tyr Glu Trp Val His Lys Pro Arg Glu Met Asp Thr Ser Arg Arg 335 340 345 Lys Phe His Leu <210> SEQ ID NO 2 <211> LENGTH: 169 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1850310 <400> SEQUENCE: 2 Met Gln Cys Leu Leu Pro Tyr Gln Ser Lys Glu Pro Ser Cys Leu 1 5 10 15 Pro Pro Leu Pro Leu Asn Leu Pro Leu Pro Pro Cys Leu Cys Pro 20 25 30 Leu Leu Gln Leu Asn Ala Ala Met Thr Arg Lys Glu Lys Thr Lys 35 40 45 Glu Gly Gln Arg Ala Ala Gln Phe Ser Ala Gly Ala Asp Ala Gly 50 55 60 Ser Gly Gly Gly Leu Ser Arg Gln Lys Asp Thr Lys Arg Pro Met 65 70 75 Leu Leu Val Ile His Asp Val Val Leu Glu Leu Leu Thr Ser Ser 80 85 90 Asp Cys His Ala Asn Pro Arg Lys Tyr Pro Thr Cys Gln Lys Ser 95 100 105 Glu Val Leu Gly Val Ser Ile Tyr Val Ser Ile Cys Pro Ser Thr 110 115 120 Arg Pro Arg Asp Lys Asn Lys Thr Lys Lys Arg Cys Gln Val Leu 125 130 135 Glu Ala Val Leu Val Ser Lys Pro Ser Gly Ser Cys His Gln Gly 140 145 150 Ser Phe Glu Ile Val Pro His Val Lys Gly Asn Leu Ala Phe Thr 155 160 165 Ser Ser Asn His <210> SEQ ID NO 3 <211> LENGTH: 316 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1887020 <400> SEQUENCE: 3 Met Glu Ser Asn Val Lys Val Gln Arg Gln Glu Gly Ala Lys Val 1 5 10 15 Ser Leu Met Ser Pro Asp Gln Leu Arg Asn Lys Phe Pro Trp Ile 20 25 30 Asn Thr Glu Gly Val Ala Leu Ala Ser Tyr Gly Met Glu Asp Glu 35 40 45 Gly Trp Phe Asp Pro Trp Cys Leu Leu Gln Gly Leu Arg Arg Lys 50 55 60 Val Gln Ser Leu Gly Val Leu Phe Cys Gln Gly Glu Val Thr Arg 65 70 75 Phe Val Ser Ser Ser Gln Arg Met Leu Thr Thr Asp Asp Lys Ala 80 85 90 Val Val Leu Lys Arg Ile His Glu Val His Val Lys Met Asp Arg 95 100 105 Ser Leu Glu Tyr Gln Pro Val Glu Cys Ala Ile Val Ile Asn Ala 110 115 120 Ala Gly Ala Trp Ser Ala Gln Ile Ala Ala Leu Ala Gly Val Gly 125 130 135 Glu Gly Pro Pro Gly Thr Leu Gln Gly Thr Lys Leu Pro Val Glu 140 145 150 Pro Arg Lys Arg Tyr Val Tyr Val Trp His Cys Pro Gln Gly Pro 155 160 165 Gly Leu Glu Thr Pro Leu Val Ala Asp Thr Ser Gly Ala Tyr Phe 170 175 180 Arg Arg Glu Gly Leu Gly Ser Asn Tyr Leu Gly Gly Arg Ser Pro 185 190 195 Thr Glu Gln Glu Glu Pro Asp Pro Ala Asn Leu Glu Val Asp His 200 205 210 Asp Phe Phe Gln Asp Lys Val Trp Pro His Leu Ala Leu Arg Val 215 220 225 Pro Ala Phe Glu Thr Leu Lys Val Gln Ser Ala Trp Ala Gly Tyr 230 235 240 Tyr Asp Tyr Asn Thr Phe Asp Gln Asn Gly Val Val Gly Pro His 245 250 255 Pro Leu Val Val Asn Met Tyr Phe Ala Thr Gly Phe Ser Gly His 260 265 270 Gly Leu Gln Gln Ala Pro Gly Ile Gly Arg Ala Val Ala Glu Met 275 280 285 Val Leu Lys Gly Arg Phe Gln Thr Ile Asp Leu Ser Pro Phe Leu 290 295 300 Phe Thr Arg Phe Tyr Leu Gly Glu Lys Ile Gln Glu Asn Asn Ile 305 310 315 Ile <210> SEQ ID NO 4 <211> LENGTH: 220 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1911421 <400> SEQUENCE: 4 Met Lys Ser Val Ile Tyr His Ala Leu Ser Gln Lys Glu Ala Asn 1 5 10 15 Asp Ser Asp Val Gln Pro Ser Gly Ala Gln Arg Ala Glu Ala Phe 20 25 30 Val Arg Ala Phe Leu Lys Arg Ser Thr Pro Arg Met Ser Pro Gln 35 40 45 Ala Arg Glu Asp Gln Leu Gln Arg Lys Ala Val Val Leu Glu Tyr 50 55 60 Phe Thr Arg His Lys Arg Lys Glu Lys Lys Lys Lys Ala Lys Gly 65 70 75 Leu Ser Ala Arg Gln Arg Arg Glu Leu Arg Leu Phe Asp Ile Lys 80 85 90 Pro Glu Gln Gln Arg Tyr Ser Leu Phe Leu Pro Leu His Glu Leu 95 100 105 Trp Lys Gln Tyr Ile Arg Asp Leu Cys Ser Gly Leu Lys Pro Asp 110 115 120 Thr Gln Pro Gln Met Ile Gln Ala Lys Leu Leu Lys Ala Asp Leu 125 130 135 His Gly Ala Ile Ile Ser Val Thr Lys Ser Lys Cys Pro Ser Tyr 140 145 150 Val Gly Ile Thr Gly Ile Leu Leu Gln Glu Thr Lys His Ile Phe 155 160 165 Lys Ile Ile Thr Lys Glu Asp Arg Leu Lys Val Ile Pro Lys Leu 170 175 180 Asn Cys Val Phe Thr Val Glu Thr Asp Gly Phe Ile Ser Tyr Ile 185 190 195 Tyr Gly Ser Lys Phe Gln Leu Arg Ser Ser Glu Arg Ser Ala Lys 200 205 210 Lys Phe Lys Ala Lys Gly Thr Ile Asp Leu 215 220 <210> SEQ ID NO 5 <211> LENGTH: 235 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: - <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1911910 <400> SEQUENCE: 5 Met Gly Ser Thr Glu Ser Ser Glu Gly Arg Arg Val Ser Phe Gly 1 5 10 15 Val Asp Glu Glu Glu Arg Val Arg Val Leu Gln Gly Val Arg Leu 20 25 30 Ser Glu Asn Val Val Asn Arg Met Lys Glu Pro Ser Ser Pro Pro 35 40 45 Pro Ala Pro Thr Ser Ser Thr Phe Gly Leu Gln Asp Gly Asn Leu 50 55 60 Arg Ala Pro His Lys Glu Ser Thr Leu Pro Arg Ser Gly Ser Ser 65 70 75 Gly Gly Gln Gln Pro Ser Gly Met Lys Glu Gly Val Lys Arg Tyr 80 85 90 Glu Gln Glu His Ala Ala Ile Gln Asp Lys Leu Phe Gln Val Ala 95 100 105 Lys Arg Glu Arg Glu Ala Ala Thr Lys His Ser Lys Ala Ser Leu 110 115 120 Pro Thr Gly Glu Gly Ser Ile Ser His Glu Glu Gln Lys Ser Val 125 130 135 Arg Leu Ala Arg Glu Leu Glu Ser Arg Glu Ala Glu Leu Arg Arg 140 145 150 Arg Asp Thr Phe Tyr Lys Glu Gln Leu Glu Arg Ile Glu Arg Lys 155 160 165 Asn Ala Glu Met Tyr Lys Leu Ser Ser Glu Gln Phe His Glu Ala 170 175 180 Ala Ser Lys Met Glu Ser Thr Ile Lys Pro Arg Arg Val Glu Pro 185 190 195 Val Cys Ser Gly Leu Gln Ala Gln Ile Leu His Cys Tyr Arg Asp 200 205 210 Arg Pro His Glu Val Leu Leu Cys Ser Asp Leu Val Lys Ala Tyr 215 220 225 Gln Arg Cys Val Ser Ala Ala His Lys Gly 230 235 <210> SEQ ID NO 6 <211> LENGTH: 487 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1928920 <400> SEQUENCE: 6 Met Ala Ser Ser Ala Glu Gly Asp Glu Gly Thr Val Val Ala Leu 1 5 10 15 Ala Gly Val Leu Gln Ser Gly Phe Gln Glu Leu Ser Leu Asn Lys 20 25 30 Leu Ala Thr Ser Leu Gly Ala Ser Glu Gln Ala Leu Arg Leu Ile 35 40 45 Ile Ser Ile Phe Leu Gly Tyr Pro Phe Ala Leu Phe Tyr Arg His 50 55 60 Tyr Leu Phe Tyr Lys Glu Thr Tyr Leu Ile His Leu Phe His Thr 65 70 75 Phe Thr Gly Leu Ser Ile Ala Tyr Phe Asn Phe Gly Asn Gln Leu 80 85 90 Tyr His Ser Leu Leu Cys Ile Val Leu Gln Phe Leu Ile Leu Arg 95 100 105 Leu Met Gly Arg Thr Ile Thr Ala Val Leu Thr Thr Phe Cys Phe 110 115 120 Gln Met Ala Tyr Leu Leu Ala Gly Tyr Tyr Tyr Thr Ala Thr Gly 125 130 135 Asn Tyr Asp Ile Lys Trp Thr Met Pro His Cys Val Leu Thr Leu 140 145 150 Lys Leu Ile Gly Leu Ala Val Asp Tyr Phe Asp Gly Gly Lys Asp 155 160 165 Gln Asn Ser Leu Ser Ser Glu Gln Gln Lys Tyr Ala Ile Arg Gly 170 175 180 Val Pro Ser Leu Leu Glu Val Ala Gly Phe Ser Tyr Phe Tyr Gly 185 190 195 Ala Phe Leu Val Gly Pro Gln Phe Ser Met Asn His Tyr Met Lys 200 205 210 Leu Val Gln Gly Glu Leu Ile Asp Ile Pro Gly Lys Ile Pro Asn 215 220 225 Ser Ile Ile Pro Ala Leu Lys Arg Leu Ser Leu Gly Leu Phe Tyr 230 235 240 Leu Val Gly Tyr Thr Leu Leu Ser Pro His Ile Thr Glu Asp Tyr 245 250 255 Leu Leu Thr Glu Asp Tyr Asp Asn His Pro Phe Trp Phe Arg Cys 260 265 270 Met Tyr Met Leu Ile Trp Gly Lys Phe Val Leu Tyr Lys Tyr Val 275 280 285 Thr Cys Trp Leu Val Thr Glu Gly Val Cys Ile Leu Thr Gly Leu 290 295 300 Gly Phe Asn Gly Phe Glu Glu Lys Gly Lys Ala Lys Trp Asp Ala 305 310 315 Cys Ala Asn Met Lys Val Trp Leu Phe Glu Thr Asn Pro Arg Phe 320 325 330 Thr Gly Thr Ile Ala Ser Phe Asn Ile Asn Thr Asn Ala Trp Val 335 340 345 Ala Arg Tyr Ile Phe Lys Arg Leu Lys Phe Leu Gly Asn Lys Glu 350 355 360 Leu Ser Gln Gly Leu Ser Leu Leu Phe Leu Ala Leu Trp His Gly 365 370 375 Leu His Ser Gly Tyr Leu Val Cys Phe Gln Met Glu Phe Leu Ile 380 385 390 Val Ile Val Glu Arg Gln Ala Ala Arg Leu Ile Gln Glu Ser Pro 395 400 405 Thr Leu Ser Lys Leu Ala Ala Ile Thr Val Leu Gln Pro Phe Tyr 410 415 420 Tyr Leu Val Gln Gln Thr Ile His Trp Leu Phe Met Gly Tyr Ser 425 430 435 Met Thr Ala Phe Cys Leu Phe Thr Trp Asp Lys Trp Leu Lys Val 440 445 450 Tyr Lys Ser Ile Tyr Phe Leu Gly His Ile Phe Phe Leu Ser Leu 455 460 465 Leu Phe Ile Leu Pro Tyr Ile His Lys Ala Met Val Pro Arg Lys 470 475 480 Glu Lys Leu Lys Lys Met Glu 485 <210> SEQ ID NO 7 <211> LENGTH: 212 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2170846 <400> SEQUENCE: 7 Met Ala Ala Pro Pro Gln Leu Arg Ala Leu Leu Val Val Val Asn 1 5 10 15 Ala Leu Leu Arg Lys Arg Arg Tyr His Ala Ala Leu Ala Val Leu 20 25 30 Lys Gly Phe Arg Asn Gly Ala Val Tyr Gly Ala Lys Ile Arg Ala 35 40 45 Pro His Ala Leu Val Met Thr Phe Leu Phe Arg Asn Gly Ser Leu 50 55 60 Gln Glu Lys Leu Trp Ala Ile Leu Gln Ala Thr Tyr Ile His Ser 65 70 75 Trp Asn Leu Ala Arg Phe Val Phe Thr Tyr Lys Gly Leu Arg Ala 80 85 90 Leu Gln Ser Tyr Ile Gln Gly Lys Thr Tyr Pro Ala His Ala Phe 95 100 105 Leu Ala Ala Phe Leu Gly Gly Ile Leu Val Phe Gly Glu Asn Asn 110 115 120 Asn Ile Asn Ser Gln Ile Asn Met Tyr Leu Leu Ser Arg Val Leu 125 130 135 Phe Ala Leu Ser Arg Leu Ala Val Glu Lys Gly Tyr Ile Pro Glu 140 145 150 Pro Arg Trp Asp Pro Phe Pro Leu Leu Thr Ala Val Val Trp Gly 155 160 165 Leu Val Leu Trp Leu Phe Glu Tyr His Arg Ser Thr Leu Gln Pro 170 175 180 Ser Leu Gln Ser Ser Met Thr Tyr Leu Tyr Glu Asp Ser Asn Val 185 190 195 Trp His Asp Ile Ser Asp Phe Leu Ile Tyr Asn Lys Ser Arg Pro 200 205 210 Ser Asn <210> SEQ ID NO 8 <211> LENGTH: 241 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2176361 <400> SEQUENCE: 8 Met Ala Pro Val Arg Arg Ser Ala Lys Trp Arg Pro Gly Gly Ile 1 5 10 15 Glu Ala Arg Gly Glu Gly Val Ser Thr Val Gly Tyr Arg Asn Lys 20 25 30 Asn Val Arg Gln Lys Thr Trp Arg Pro Asn His Pro Gln Ala Phe 35 40 45 Val Gly Ser Val Arg Glu Gly Gln Gly Phe Ala Phe Arg Arg Lys 50 55 60 Leu Lys Ile Gln Gln Ser Tyr Lys Lys Leu Leu Arg Lys Glu Lys 65 70 75 Lys Ala Gln Thr Ser Leu Glu Ser Gln Phe Thr Asp Arg Tyr Pro 80 85 90 Asp Asn Leu Lys His Leu Tyr Leu Ala Glu Glu Glu Arg His Arg 95 100 105 Lys Gln Ala Arg Lys Val Asp His Pro Leu Ser Glu Gln Val His 110 115 120 Gln Pro Leu Leu Glu Glu Gln Cys Ser Ile Asp Glu Pro Leu Phe 125 130 135 Glu Asp Gln Cys Ser Phe Asp Gln Pro Gln Pro Glu Glu Gln Cys 140 145 150 Ile Lys Thr Val Asn Ser Phe Thr Ile Pro Lys Lys Asn Lys Lys 155 160 165 Lys Thr Ser Asn Gln Lys Ala Gln Glu Glu Tyr Glu Gln Ile Gln 170 175 180 Ala Lys Arg Ala Ala Lys Lys Gln Glu Phe Glu Arg Arg Lys Gln 185 190 195 Glu Arg Glu Glu Ala Gln Arg Gln Tyr Lys Lys Lys Lys Met Glu 200 205 210 Val Phe Lys Ile Leu Asn Lys Lys Thr Lys Lys Gly Gln Pro Asn 215 220 225 Leu Asn Val Gln Met Glu Tyr Leu Leu Gln Lys Ile Gln Glu Lys 230 235 240 Cys <210> SEQ ID NO 9 <211> LENGTH: 375 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2212732 <400> SEQUENCE: 9 Met Pro Gln Glu Leu Pro Gln Ser Pro Arg Thr Arg Gln Pro Glu 1 5 10 15 Pro Asp Phe Tyr Cys Val Lys Trp Ile Pro Trp Lys Gly Glu Gln 20 25 30 Thr Pro Ile Ile Thr Gln Ser Thr Asn Gly Pro Cys Pro Leu Leu 35 40 45 Ala Ile Met Asn Ile Leu Phe Leu Gln Trp Lys Val Lys Leu Pro 50 55 60 Pro Gln Lys Glu Val Ile Thr Ser Asp Glu Leu Met Ala His Leu 65 70 75 Gly Asn Cys Leu Leu Ser Ile Lys Pro Gln Glu Lys Ser Glu Gly 80 85 90 Leu Gln Leu Asn Phe Gln Gln Asn Val Asp Asp Ala Met Thr Val 95 100 105 Leu Pro Lys Leu Ala Thr Gly Leu Asp Val Asn Val Arg Phe Thr 110 115 120 Gly Val Ser Asp Phe Glu Tyr Thr Pro Glu Cys Ser Val Phe Asp 125 130 135 Leu Leu Gly Ile Pro Leu Tyr His Gly Trp Leu Val Asp Pro Gln 140 145 150 Gln Ser Pro Glu Ala Val Arg Ala Val Gly Lys Leu Ser Tyr Asn 155 160 165 Gln Leu Val Glu Arg Ile Ile Thr Cys Lys His Ser Ser Asp Thr 170 175 180 Asn Leu Val Thr Glu Gly Leu Ile Ala Glu Gln Phe Leu Glu Thr 185 190 195 Thr Ala Ala Gln Leu Thr Tyr His Gly Leu Cys Glu Leu Thr Ala 200 205 210 Ala Ala Lys Glu Gly Glu Leu Ser Val Phe Phe Arg Asn Asn His 215 220 225 Phe Ser Thr Met Thr Lys His Lys Ser His Leu Tyr Leu Leu Val 230 235 240 Thr Asp Gln Gly Phe Leu Gln Glu Glu Gln Val Val Trp Glu Ser 245 250 255 Leu His Asn Val Asp Gly Asp Ser Cys Phe Cys Asp Ser Asp Phe 260 265 270 His Leu Ser His Ser Leu Gly Lys Gly Pro Gly Ala Glu Gly Gly 275 280 285 Ser Gly Ser Pro Glu Lys Gln Leu Gln Val Asp Gln Asp Tyr Leu 290 295 300 Ile Ala Leu Ser Leu Gln Gln Gln Gln Pro Arg Gly Pro Leu Gly 305 310 315 Leu Thr Asp Leu Glu Leu Ala Gln Gln Leu Gln Gln Glu Glu Tyr 320 325 330 Gln Gln Gln Gln Ala Ala Gln Pro Val Arg Met Arg Thr Arg Val 335 340 345 Leu Ser Leu Gln Gly Arg Gly Ala Thr Ser Gly Arg Pro Ala Gly 350 355 360 Glu Arg Arg Gln Arg Pro Lys His Glu Ser Asp Cys Ile Leu Leu 365 370 375 <210> SEQ ID NO 10 <211> LENGTH: 429 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2303457 <400> SEQUENCE: 10 Met Ser Asn Arg Asn Asn Asn Lys Leu Pro Ser Asn Leu Pro Gln 1 5 10 15 Leu Gln Asn Leu Ile Lys Arg Asp Pro Pro Ala Tyr Ile Glu Glu 20 25 30 Phe Leu Gln Gln Tyr Asn His Tyr Lys Ser Asn Val Glu Ile Phe 35 40 45 Lys Leu Gln Pro Asn Lys Pro Ser Lys Glu Leu Ala Glu Leu Val 50 55 60 Met Phe Met Ala Gln Ile Ser His Cys Tyr Pro Glu Tyr Leu Ser 65 70 75 Asn Phe Pro Gln Glu Val Lys Asp Leu Leu Ser Cys Asn His Thr 80 85 90 Val Leu Asp Pro Asp Leu Arg Met Thr Phe Cys Lys Ala Leu Ile 95 100 105 Leu Leu Arg Asn Lys Asn Leu Ile Asn Pro Ser Ser Leu Leu Glu 110 115 120 Leu Phe Phe Glu Leu Phe Arg Cys His Asp Lys Leu Leu Arg Lys 125 130 135 Thr Leu Tyr Thr His Ile Val Thr Asp Ile Lys Asn Ile Asn Ala 140 145 150 Lys His Lys Asn Asn Lys Val Asn Val Val Leu Gln Asn Phe Met 155 160 165 Tyr Thr Met Leu Arg Asp Ser Asn Ala Thr Ala Ala Lys Met Ser 170 175 180 Leu Asp Val Met Ile Glu Leu Tyr Arg Arg Asn Ile Trp Asn Asp 185 190 195 Ala Lys Thr Val Asn Val Ile Thr Thr Ala Cys Phe Ser Lys Val 200 205 210 Thr Lys Ile Leu Val Ala Ala Leu Thr Phe Phe Leu Gly Lys Asp 215 220 225 Glu Asp Glu Lys Gln Asp Ser Asp Ser Glu Ser Glu Asp Asp Gly 230 235 240 Pro Thr Ala Arg Asp Leu Leu Val Gln Tyr Ala Thr Gly Lys Lys 245 250 255 Ser Ser Lys Asn Lys Lys Lys Leu Glu Lys Ala Met Lys Val Leu 260 265 270 Lys Lys Gln Lys Lys Lys Lys Lys Pro Glu Val Phe Asn Phe Ser 275 280 285 Ala Ile His Leu Ile His Asp Pro Gln Asp Phe Ala Glu Lys Leu 290 295 300 Leu Lys Gln Leu Glu Cys Cys Lys Glu Arg Phe Glu Val Lys Met 305 310 315 Met Leu Met Asn Leu Ile Ser Arg Leu Val Gly Ile His Glu Leu 320 325 330 Phe Leu Phe Asn Phe Tyr Pro Phe Leu Lys Arg Phe Leu Lys Pro 335 340 345 His Gln Arg Glu Val Thr Lys Ile Leu Leu Phe Val Glu Lys Asp 350 355 360 Ser His His Leu Val Pro Gln Gly Phe Phe Asn Ser Trp Leu Met 365 370 375 Leu Gly Glu Lys Ile Phe Phe Asn Gly Lys Lys Ser Gly Lys Met 380 385 390 Leu Met Thr Val Gly Asn Leu Met Val Lys Arg Gly Val Tyr Lys 395 400 405 Arg Ser Lys Val Phe Leu Gly Gly Asn Ser Val Gly Arg Asn Phe 410 415 420 Phe Gln Lys Asn Pro Gly Gly Ser Ser 425 <210> SEQ ID NO 11 <211> LENGTH: 329 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2317552 <400> SEQUENCE: 11 Met Glu Val Ala Glu Pro Ser Ser Pro Thr Glu Glu Glu Glu Glu 1 5 10 15 Glu Glu Glu His Ser Ala Glu Pro Arg Pro Arg Thr Arg Ser Asn 20 25 30 Pro Glu Gly Ala Glu Asp Arg Ala Val Gly Ala Gln Ala Ser Val 35 40 45 Gly Ser Arg Ser Glu Gly Glu Gly Glu Ala Ala Ser Ala Asp Asp 50 55 60 Gly Ser Leu Asn Thr Ser Gly Ala Gly Pro Lys Ser Trp Gln Val 65 70 75 Pro Pro Pro Ala Pro Glu Val Gln Ile Arg Thr Pro Arg Val Asn 80 85 90 Cys Pro Glu Lys Val Ile Ile Cys Leu Asp Leu Ser Glu Glu Met 95 100 105 Ser Leu Pro Lys Leu Glu Ser Phe Asn Gly Ser Lys Thr Asn Ala 110 115 120 Leu Asn Val Ser Gln Lys Met Ile Glu Met Phe Val Arg Thr Lys 125 130 135 His Lys Ile Asp Lys Ser His Glu Phe Ala Leu Val Val Val Asn 140 145 150 Asp Asp Thr Ala Trp Leu Ser Gly Leu Thr Ser Asp Pro Arg Glu 155 160 165 Leu Cys Ser Cys Leu Tyr Asp Leu Glu Thr Ala Ser Cys Ser Thr 170 175 180 Phe Asn Leu Glu Gly Leu Phe Ser Leu Ile Gln Gln Lys Thr Glu 185 190 195 Leu Pro Val Thr Glu Asn Val Gln Thr Ile Pro Pro Pro Tyr Val 200 205 210 Val Arg Thr Ile Leu Val Tyr Ser Arg Pro Pro Cys Gln Pro Gln 215 220 225 Phe Ser Leu Thr Glu Pro Met Lys Lys Met Phe Gln Cys Pro Tyr 230 235 240 Phe Phe Phe Asp Val Val Tyr Ile His Asn Gly Thr Glu Glu Lys 245 250 255 Glu Glu Glu Met Ser Trp Lys Asp Met Phe Ala Phe Met Gly Ser 260 265 270 Leu Asp Thr Lys Gly Thr Ser Tyr Lys Tyr Glu Val Ala Leu Ala 275 280 285 Gly Pro Ala Leu Glu Leu His Asn Cys Met Ala Lys Leu Leu Ala 290 295 300 His Pro Leu Gln Arg Pro Cys Gln Ser His Ala Ser Tyr Ser Leu 305 310 315 Leu Glu Glu Glu Asp Glu Ala Ile Glu Val Glu Ala Thr Val 320 325 <210> SEQ ID NO 12 <211> LENGTH: 476 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2416366 <400> SEQUENCE: 12 Met Gln Asn Asp Ser Phe His Ser Asp Ser His Met Asp Arg Lys 1 5 10 15 Lys Phe His Ser Ser Asp Ser Glu Glu Glu Glu His Lys Lys Gln 20 25 30 Lys Met Asp Ser Asp Glu Asp Glu Lys Glu Gly Glu Glu Glu Lys 35 40 45 Val Ala Lys Arg Lys Ala Ala Val Leu Ser Asp Ser Glu Asp Glu 50 55 60 Glu Lys Ala Ser Ala Lys Lys Ser Arg Val Val Ser Asp Ala Asp 65 70 75 Asp Ser Asp Ser Asp Ala Val Ser Asp Lys Ser Gly Lys Arg Glu 80 85 90 Lys Thr Ile Ala Ser Asp Ser Glu Glu Glu Ala Gly Lys Glu Leu 95 100 105 Ser Asp Lys Lys Asn Glu Glu Lys Asp Leu Phe Gly Ser Asp Ser 110 115 120 Glu Ser Gly Asn Glu Glu Glu Asn Leu Ile Ala Asp Ile Phe Gly 125 130 135 Glu Ser Gly Asp Glu Glu Glu Glu Glu Phe Thr Gly Phe Asn Gln 140 145 150 Glu Asp Leu Glu Glu Glu Lys Gly Glu Thr Gln Val Lys Glu Ala 155 160 165 Glu Asp Ser Asp Ser Asp Asp Asn Ile Lys Arg Gly Lys His Met 170 175 180 Asp Phe Leu Ser Asp Phe Glu Met Met Leu Gln Arg Lys Lys Ser 185 190 195 Met Ser Gly Lys Arg Arg Arg Asn Arg Asp Gly Gly Thr Phe Ile 200 205 210 Ser Asp Ala Asp Asp Val Val Ser Ala Met Ile Val Lys Met Asn 215 220 225 Glu Ala Ala Glu Glu Asp Arg Gln Leu Asn Asn Gln Lys Lys Pro 230 235 240 Ala Leu Lys Lys Leu Thr Leu Leu Pro Ala Val Val Met His Leu 245 250 255 Lys Lys Gln Asp Leu Lys Glu Thr Phe Ile Asp Ser Gly Val Met 260 265 270 Ser Ala Ile Lys Glu Trp Leu Ser Pro Leu Pro Asp Arg Ser Leu 275 280 285 Pro Ala Leu Lys Ile Arg Glu Glu Leu Leu Lys Ile Leu Gln Glu 290 295 300 Leu Pro Ser Val Ser Gln Glu Thr Leu Lys His Ser Gly Ile Gly 305 310 315 Arg Ala Val Met Tyr Leu Tyr Lys His Pro Lys Glu Ser Arg Ser 320 325 330 Asn Lys Asp Met Ala Gly Lys Leu Ile Asn Glu Trp Ser Arg Pro 335 340 345 Ile Phe Gly Leu Thr Ser Asn Tyr Lys Gly Met Thr Arg Glu Glu 350 355 360 Arg Glu Gln Arg Asp Leu Glu Gln Met Pro Gln Arg Arg Arg Met 365 370 375 Asn Ser Thr Gly Gly Gln Thr Pro Arg Arg Asp Leu Glu Lys Val 380 385 390 Leu Thr Gly Glu Glu Lys Ala Leu Arg Pro Gly Asp Pro Gly Phe 395 400 405 Cys Ala Arg Ala Arg Val Pro Met Pro Ser Asn Lys Asp Tyr Val 410 415 420 Val Arg Pro Lys Trp Asn Val Glu Met Glu Ser Ser Arg Phe Gln 425 430 435 Ala Thr Ser Lys Lys Gly Ile Ser Arg Leu Asp Lys Gln Met Arg 440 445 450 Lys Phe Thr Asp Ile Arg Lys Lys Ser Arg Ser Ala His Ala Val 455 460 465 Lys Ile Ser Ile Glu Gly Asn Lys Met Pro Leu 470 475 <210> SEQ ID NO 13 <211> LENGTH: 366 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2472980 <400> SEQUENCE: 13 Met Ala Ala Ala Tyr Phe Pro Asp Cys Ile Val Arg Pro Phe Gly 1 5 10 15 Ser Ser Val Asn Thr Phe Gly Lys Leu Gly Cys Asp Leu Asp Met 20 25 30 Phe Leu Asp Leu Asp Glu Thr Arg Asn Leu Ser Ala His Lys Ile 35 40 45 Ser Gly Asn Phe Leu Met Glu Phe Gln Val Lys Asn Val Pro Ser 50 55 60 Glu Arg Ile Ala Thr Gln Lys Ile Leu Ser Val Leu Gly Glu Cys 65 70 75 Leu Asp His Phe Gly Pro Gly Cys Val Gly Val Gln Lys Ile Leu 80 85 90 Asn Ala Arg Cys Pro Leu Val Arg Phe Ser His Gln Ala Ser Gly 95 100 105 Phe Gln Cys Asp Leu Thr Thr Asn Asn Arg Ile Ala Leu Thr Ser 110 115 120 Ser Glu Leu Leu Tyr Ile Tyr Gly Ala Leu Asp Ser Arg Val Arg 125 130 135 Ala Leu Val Phe Ser Val Arg Cys Trp Ala Arg Ala His Ser Leu 140 145 150 Thr Ser Ser Ile Pro Gly Ala Trp Ile Thr Asn Phe Ser Leu Thr 155 160 165 Met Met Val Ile Phe Phe Leu Gln Arg Arg Ser Pro Pro Ile Leu 170 175 180 Pro Thr Leu Asp Ser Leu Lys Thr Leu Ala Asp Ala Glu Asp Lys 185 190 195 Cys Val Ile Glu Gly Asn Asn Cys Thr Phe Val Arg Asp Leu Ser 200 205 210 Arg Ile Lys Pro Ser Gln Asn Thr Glu Thr Leu Glu Leu Leu Leu 215 220 225 Lys Glu Phe Phe Glu Tyr Phe Gly Asn Phe Ala Phe Asp Lys Asn 230 235 240 Ser Ile Asn Ile Arg Gln Gly Arg Glu Gln Asn Lys Pro Asp Ser 245 250 255 Ser Pro Leu Tyr Ile Gln Asn Pro Phe Glu Thr Ser Leu Asn Ile 260 265 270 Ser Lys Asn Val Ser Gln Ser Gln Leu Gln Lys Phe Val Asp Leu 275 280 285 Ala Arg Glu Ser Ala Trp Ile Leu Gln Gln Glu Asp Thr Asp Arg 290 295 300 Pro Ser Ile Ser Ser Asn Arg Pro Trp Gly Leu Val Ser Leu Leu 305 310 315 Leu Pro Ser Ala Pro Asn Arg Lys Ser Phe Thr Lys Lys Lys Ser 320 325 330 Asn Lys Phe Ala Ile Glu Thr Val Lys Asn Leu Leu Glu Ser Leu 335 340 345 Lys Gly Asn Arg Thr Glu Asn Phe Thr Lys Thr Ser Gly Lys Arg 350 355 360 Thr Ile Ser Thr Gln Thr 365 <210> SEQ ID NO 14 <211> LENGTH: 152 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2541640 <400> SEQUENCE: 14 Met Gly Gly Val Gly Val Ala Glu Ala Ala Arg Pro Leu Leu Ser 1 5 10 15 Trp Pro Thr Ile Ser Leu Thr Ile Phe Thr Ala Val Asn Ser Ser 20 25 30 Gln Gly Gly Gly Leu Val Gln Arg Gln Leu Arg Phe His Asn Ser 35 40 45 His Arg Val Leu Cys Arg Arg Cys Pro Cys Pro Pro Thr Pro Ala 50 55 60 Trp Trp Glu Cys Asp Ala Arg Leu Leu Pro Pro Pro Trp Pro Pro 65 70 75 Val Pro Pro Ala Ser Thr Ser Pro Glu Ile Leu Pro Thr Pro His 80 85 90 Leu His Arg Ser Pro His Ala Pro Gly Ala Pro Lys Pro Pro Pro 95 100 105 Asn Pro Thr His Pro Gly Ala Gly Thr Gly Val Ser Glu Leu Ser 110 115 120 Gln Gly Pro Trp Glu Val Ala Gly Thr Gly Ala Ser Cys Ser Leu 125 130 135 Phe His Phe Pro Phe Arg Ile Trp Pro Gly Trp Arg Thr Gly Gln 140 145 150 Asp Gly <210> SEQ ID NO 15 <211> LENGTH: 233 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2695204 <400> SEQUENCE: 15 Met Gly Arg Arg Leu Lys Gly Ala Arg Arg Leu Lys Leu Ser Pro 1 5 10 15 Leu Arg Ser Leu Arg Lys Gly Pro Gly Leu Leu Ser Pro Pro Ser 20 25 30 Ala Ser Pro Val Pro Thr Pro Ala Val Ser Arg Thr Leu Leu Gly 35 40 45 Asn Phe Glu Glu Ser Leu Leu Arg Gly Arg Phe Ala Pro Ser Gly 50 55 60 His Ile Glu Gly Phe Thr Ala Glu Ile Gly Ala Ser Gly Ser Tyr 65 70 75 Cys Pro Gln His Val Thr Leu Pro Val Thr Val Thr Phe Phe Asp 80 85 90 Val Ser Glu Gln Asn Ala Pro Ala Pro Phe Leu Gly Ile Val Asp 95 100 105 Leu Asn Pro Leu Gly Arg Lys Gly Tyr Ser Val Pro Lys Val Gly 110 115 120 Thr Val Gln Val Thr Leu Phe Asn Pro Asn Gln Thr Val Val Lys 125 130 135 Met Phe Leu Val Thr Phe Asp Phe Ser Asp Met Pro Ala Ala His 140 145 150 Met Thr Phe Leu Arg His Arg Leu Phe Leu Val Pro Val Gly Glu 155 160 165 Glu Gly Asn Ala Asn Pro Thr His Arg Leu Leu Cys Tyr Leu Leu 170 175 180 His Leu Arg Phe Arg Ser Ser Arg Ser Gly Arg Leu Ser Leu His 185 190 195 Gly Asp Ile Arg Leu Leu Phe Ser Arg Arg Ser Leu Glu Leu Asp 200 205 210 Thr Gly Leu Pro Tyr Glu Leu Gln Ala Val Thr Glu Ala Pro His 215 220 225 Asn Pro Arg Tyr Ser Pro Leu Pro 230 <210> SEQ ID NO 16 <211> LENGTH: 357 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2805526 <400> SEQUENCE: 16 Met Glu Val Leu Arg Pro Gln Leu Ile Arg Ile Asp Gly Arg Asn 1 5 10 15 Tyr Arg Lys Asn Pro Val Gln Glu Gln Thr Tyr Gln His Glu Glu 20 25 30 Asp Glu Glu Asp Phe Tyr Gln Gly Ser Met Glu Cys Ala Asp Glu 35 40 45 Pro Cys Asp Ala Tyr Glu Val Glu Gln Thr Pro Gln Gly Phe Arg 50 55 60 Ser Thr Leu Arg Ala Pro Ser Leu Leu Tyr Lys His Ile Val Gly 65 70 75 Lys Arg Gly Asp Thr Arg Lys Lys Ile Glu Met Glu Thr Lys Thr 80 85 90 Ser Ile Ser Ile Pro Lys Pro Gly Gln Asp Gly Glu Ile Val Ile 95 100 105 Thr Gly Gln His Arg Asn Gly Val Ile Ser Ala Arg Thr Arg Ile 110 115 120 Asp Val Leu Leu Asp Thr Phe Arg Arg Lys Gln Pro Phe Thr His 125 130 135 Phe Leu Ala Phe Phe Leu Asn Glu Val Glu Val Gln Glu Gly Phe 140 145 150 Leu Arg Phe Gln Glu Glu Val Leu Ala Lys Cys Ser Met Asp His 155 160 165 Gly Val Asp Ser Ser Ile Phe Gln Asn Pro Lys Lys Leu His Leu 170 175 180 Thr Ile Gly Met Leu Val Leu Leu Ser Glu Glu Glu Ile Gln Gln 185 190 195 Thr Cys Glu Met Leu Gln Gln Cys Lys Glu Glu Phe Ile Asn Asp 200 205 210 Ile Ser Gly Gly Lys Pro Leu Glu Val Glu Met Ala Gly Ile Glu 215 220 225 Tyr Met Asn Asp Asp Pro Gly Met Val Asp Val Leu Tyr Ala Lys 230 235 240 Val His Met Lys Asp Gly Ser Asn Arg Leu Gln Glu Leu Val Asp 245 250 255 Arg Val Leu Glu Arg Phe Gln Ala Ser Gly Leu Ile Val Lys Glu 260 265 270 Trp Asn Ser Val Lys Leu His Ala Thr Val Met Asn Thr Leu Phe 275 280 285 Arg Lys Asp Pro Asn Ala Glu Gly Arg Tyr Asn Leu Tyr Thr Ala 290 295 300 Glu Gly Lys Tyr Ile Phe Lys Glu Arg Glu Ser Phe Asp Gly Arg 305 310 315 Asn Ile Leu Lys Leu Phe Glu Asn Phe Tyr Phe Gly Ser Leu Lys 320 325 330 Leu Asn Ser Ile His Ile Ser Gln Arg Phe Thr Val Asp Ser Phe 335 340 345 Gly Asn Tyr Ala Ser Cys Gly Gln Ile Asp Phe Ser 350 355 <210> SEQ ID NO 17 <211> LENGTH: 251 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2850382 <400> SEQUENCE: 17 Met Glu Pro Gly Glu Glu Leu Glu Glu Glu Gly Ser Pro Gly Gly 1 5 10 15 Arg Glu Asp Gly Phe Thr Ala Glu His Leu Ala Ala Glu Ala Met 20 25 30 Ala Ala Asp Met Asp Pro Trp Leu Val Phe Asp Ala Arg Thr Thr 35 40 45 Pro Ala Thr Glu Leu Asp Ala Trp Leu Ala Lys Tyr Pro Pro Ser 50 55 60 Gln Val Thr Arg Tyr Gly Asp Pro Gly Ser Pro Asn Ser Glu Pro 65 70 75 Val Gly Trp Ile Ala Val Tyr Gly Gln Gly Tyr Ser Pro Asn Ser 80 85 90 Gly Asp Val Gln Gly Leu Gln Ala Ala Trp Glu Ala Leu Gln Thr 95 100 105 Ser Gly Arg Pro Ile Thr Pro Gly Thr Leu Arg Gln Leu Ala Ile 110 115 120 Thr His His Val Leu Ser Gly Lys Trp Leu Met His Leu Ala Pro 125 130 135 Gly Phe Lys Leu Asp His Ala Trp Ala Gly Ile Ala Arg Ala Val 140 145 150 Val Glu Gly Arg Leu Gln Val Ala Lys Val Ser Pro Arg Ala Lys 155 160 165 Glu Gly Gly Arg Gln Val Ile Cys Val Tyr Thr Asp Asp Phe Thr 170 175 180 Asp Arg Leu Gly Val Leu Glu Ala Asp Ser Ala Ile Arg Ala Ala 185 190 195 Gly Ile Lys Cys Leu Leu Thr Tyr Lys Pro Asp Val Tyr Thr Tyr 200 205 210 Leu Gly Ile Tyr Arg Ala Asn Arg Trp His Leu Cys Pro Thr Leu 215 220 225 Tyr Glu Ser Arg Phe Gln Leu Gly Gly Ser Ala Arg Gly Ser Arg 230 235 240 Val Leu Asp Arg Ala Asn Asn Val Glu Leu Thr 245 250 <210> SEQ ID NO 18 <211> LENGTH: 105 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2929276 <400> SEQUENCE: 18 Met Ser Ile Tyr Phe Pro Ile His Cys Pro Asp Tyr Leu Arg Ser 1 5 10 15 Ala Lys Met Thr Glu Val Met Met Asn Thr Gln Pro Met Glu Glu 20 25 30 Ile Gly Leu Ser Pro Arg Lys Asp Gly Leu Ser Tyr Gln Ile Phe 35 40 45 Pro Asp Pro Ser Asp Phe Asp Arg Cys Cys Lys Leu Lys Asp Arg 50 55 60 Leu Pro Ser Ile Val Val Glu Pro Thr Glu Gly Glu Val Glu Ser 65 70 75 Gly Glu Leu Arg Trp Pro Pro Glu Glu Phe Leu Val Gln Glu Asp 80 85 90 Glu Gln Asp Asn Cys Glu Glu Thr Ala Lys Glu Asn Lys Glu Gln 95 100 105 <210> SEQ ID NO 19 <211> LENGTH: 876 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 3033039 <400> SEQUENCE: 19 Met Thr Met Asp Ala Leu Leu Ala Arg Leu Lys Leu Leu Asn Pro 1 5 10 15 Asp Asp Leu Arg Glu Glu Ile Val Lys Ala Gly Leu Lys Cys Gly 20 25 30 Pro Ile Thr Ser Thr Thr Arg Phe Ile Phe Glu Lys Lys Leu Ala 35 40 45 Gln Ala Leu Leu Glu Gln Gly Gly Arg Leu Ser Ser Phe Tyr His 50 55 60 His Glu Ala Gly Val Thr Ala Leu Ser Gln Asp Pro Gln Arg Ile 65 70 75 Leu Lys Pro Ala Glu Gly Asn Pro Thr Asp Gln Ala Gly Phe Ser 80 85 90 Glu Asp Arg Asp Phe Gly Tyr Ser Val Gly Leu Asn Pro Pro Glu 95 100 105 Glu Glu Ala Val Thr Ser Lys Thr Cys Ser Val Pro Pro Ser Asp 110 115 120 Thr Asp Thr Tyr Arg Ala Gly Ala Thr Ala Ser Lys Glu Pro Pro 125 130 135 Leu Tyr Tyr Gly Val Cys Pro Val Tyr Glu Asp Val Pro Ala Arg 140 145 150 Asn Glu Arg Ile Tyr Val Tyr Glu Asn Lys Lys Glu Ala Leu Gln 155 160 165 Ala Val Lys Met Ile Lys Gly Ser Arg Phe Lys Ala Phe Ser Thr 170 175 180 Arg Glu Asp Ala Glu Lys Phe Ala Arg Gly Ile Cys Asp Tyr Phe 185 190 195 Pro Ser Pro Ser Lys Thr Ser Leu Pro Leu Ser Pro Val Lys Thr 200 205 210 Ala Pro Leu Phe Ser Asn Asp Arg Leu Lys Asp Gly Leu Cys Leu 215 220 225 Ser Glu Ser Glu Thr Val Asn Lys Glu Arg Ala Asn Ser Tyr Lys 230 235 240 Asn Pro Arg Thr Gln Asp Leu Thr Ala Lys Leu Arg Lys Ala Val 245 250 255 Glu Lys Gly Glu Glu Asp Thr Phe Ser Asp Leu Ile Trp Ser Asn 260 265 270 Pro Arg Tyr Leu Ile Gly Ser Gly Asp Asn Pro Thr Ile Val Gln 275 280 285 Glu Gly Cys Arg Tyr Asn Val Met His Val Ala Ala Lys Glu Asn 290 295 300 Gln Ala Ser Ile Cys Gln Leu Thr Leu Asp Val Leu Glu Asn Pro 305 310 315 Asp Phe Met Arg Leu Met Tyr Pro Asp Asp Asp Glu Ala Met Leu 320 325 330 Gln Lys Arg Ile Arg Tyr Val Val Asp Leu Tyr Leu Asn Thr Pro 335 340 345 Asp Lys Met Gly Tyr Asp Thr Pro Leu His Phe Ala Cys Lys Phe 350 355 360 Gly Asn Ala Asp Val Val Asn Val Leu Ser Ser His His Leu Ile 365 370 375 Val Lys Asn Ser Arg Asn Lys Tyr Asp Lys Thr Pro Glu Asp Val 380 385 390 Ile Cys Glu Arg Ser Lys Asn Lys Ser Val Glu Leu Lys Glu Arg 395 400 405 Ile Arg Glu Tyr Leu Lys Gly His Tyr Tyr Val Pro Leu Leu Arg 410 415 420 Ala Glu Glu Thr Ser Ser Pro Val Ile Gly Glu Leu Trp Ser Pro 425 430 435 Asp Gln Thr Ala Glu Ala Ser His Val Ser Arg Tyr Gly Gly Ser 440 445 450 Pro Arg Asp Pro Val Leu Thr Leu Arg Ala Phe Ala Gly Pro Leu 455 460 465 Ser Pro Ala Lys Ala Glu Asp Phe Arg Lys Leu Trp Lys Thr Pro 470 475 480 Pro Arg Glu Lys Ala Gly Phe Leu His His Val Lys Lys Ser Asp 485 490 495 Pro Glu Arg Gly Phe Glu Arg Val Gly Arg Glu Leu Ala His Glu 500 505 510 Leu Gly Tyr Pro Trp Val Glu Tyr Trp Glu Phe Leu Gly Cys Phe 515 520 525 Val Asp Leu Ser Ser Gln Glu Gly Leu Gln Arg Leu Glu Glu Tyr 530 535 540 Leu Thr Gln Gln Glu Ile Gly Lys Lys Ala Gln Gln Glu Thr Gly 545 550 555 Glu Arg Glu Ala Ser Cys Arg Asp Lys Ala Thr Thr Ser Gly Ser 560 565 570 Asn Ser Ile Ser Val Arg Ala Phe Leu Asp Glu Asp Asp Met Ser 575 580 585 Leu Glu Glu Ile Lys Asn Arg Gln Asn Ala Ala Arg Asn Asn Ser 590 595 600 Pro Pro Thr Val Gly Ala Phe Gly His Thr Arg Cys Ser Ala Phe 605 610 615 Pro Leu Glu Gln Glu Ala Asp Leu Ile Glu Ala Ala Glu Pro Gly 620 625 630 Gly Pro His Ser Ser Arg Asn Gly Leu Cys His Pro Leu Asn His 635 640 645 Ser Arg Thr Leu Ala Gly Lys Arg Pro Lys Ala Pro Arg Gly Glu 650 655 660 Glu Ala His Leu Pro Pro Val Ser Asp Leu Thr Val Glu Phe Asp 665 670 675 Lys Leu Asn Leu Gln Asn Ile Gly Arg Ser Val Ser Lys Thr Pro 680 685 690 Asp Glu Ser Thr Lys Thr Lys Asp Gln Ile Leu Thr Ser Arg Ile 695 700 705 Asn Ala Val Glu Arg Asp Leu Leu Glu Pro Ser Pro Ala Asp Gln 710 715 720 Leu Gly Asn Gly His Arg Arg Thr Glu Ser Glu Met Ser Ala Arg 725 730 735 Ile Ala Lys Met Ser Leu Ser Pro Ser Ser Pro Arg His Glu Asp 740 745 750 Gln Leu Glu Val Thr Arg Glu Pro Ala Arg Arg Leu Phe Leu Phe 755 760 765 Gly Glu Glu Pro Ser Lys Leu Asp Gln Asp Val Leu Ala Ala Leu 770 775 780 Glu Cys Ala Asp Val Asp Pro His Gln Phe Pro Ala Val His Arg 785 790 795 Trp Lys Ser Ala Val Leu Cys Tyr Ser Pro Ser Asp Arg Gln Ser 800 805 810 Trp Pro Ser Pro Ala Val Lys Gly Arg Phe Lys Ser Gln Leu Pro 815 820 825 Asp Leu Ser Gly Pro His Ser Tyr Ser Pro Gly Arg Asn Ser Val 830 835 840 Ala Gly Ser Asn Pro Ala Lys Pro Gly Leu Gly Ser Pro Gly Arg 845 850 855 Tyr Ser Pro Val His Gly Ser Gln Leu Arg Arg Met Ala Arg Leu 860 865 870 Ala Glu Leu Ala Ala Leu 875 <210> SEQ ID NO 20 <211> LENGTH: 505 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 3039890 <400> SEQUENCE: 20 Met Ser Arg Ser Tyr Asn Asp Glu Leu Gln Phe Leu Glu Lys Ile 1 5 10 15 Asn Lys Asn Cys Trp Arg Ile Lys Lys Gly Phe Val Pro Asn Met 20 25 30 Gln Val Glu Gly Val Phe Tyr Val Asn Asp Ala Leu Glu Lys Leu 35 40 45 Met Phe Glu Glu Leu Arg Asn Ala Cys Arg Gly Gly Gly Val Gly 50 55 60 Gly Phe Leu Pro Ala Met Lys Gln Ile Gly Asn Val Ala Ala Leu 65 70 75 Pro Gly Ile Val His Arg Ser Ile Gly Leu Pro Asp Val His Ser 80 85 90 Gly Tyr Gly Phe Ala Ile Gly Asn Met Ala Ala Phe Asp Met Asn 95 100 105 Asp Pro Glu Ala Val Val Ser Pro Gly Gly Val Gly Phe Asp Ile 110 115 120 Asn Cys Gly Val Arg Leu Leu Arg Thr Asn Leu Asp Glu Ser Asp 125 130 135 Val Gln Pro Val Lys Glu Gln Leu Ala Gln Ala Met Phe Asp His 140 145 150 Ile Pro Val Gly Val Gly Ser Lys Gly Val Ile Pro Met Asn Ala 155 160 165 Lys Asp Leu Glu Glu Ala Leu Glu Met Gly Val Asp Trp Ser Leu 170 175 180 Arg Glu Gly Tyr Ala Trp Ala Glu Asp Lys Glu His Cys Glu Glu 185 190 195 Tyr Gly Arg Met Leu Gln Ala Asp Pro Asn Lys Val Ser Ala Arg 200 205 210 Ala Lys Lys Arg Gly Leu Pro Gln Leu Gly Thr Leu Gly Ala Gly 215 220 225 Asn His Tyr Ala Glu Ile Gln Val Val Asp Glu Ile Phe Asn Glu 230 235 240 Tyr Ala Ala Lys Lys Met Gly Ile Asp His Lys Gly Gln Val Cys 245 250 255 Val Met Ile His Ser Gly Ser Arg Gly Leu Gly His Gln Val Ala 260 265 270 Thr Asp Ala Leu Val Ala Met Glu Lys Ala Met Lys Arg Asp Lys 275 280 285 Ile Ile Val Asn Asp Arg Gln Leu Ala Cys Ala Arg Ile Ala Ser 290 295 300 Pro Glu Gly Gln Asp Tyr Leu Lys Gly Met Ala Ala Ala Gly Asn 305 310 315 Tyr Ala Trp Val Asn Arg Ser Ser Met Thr Phe Leu Thr Arg Gln 320 325 330 Ala Phe Ala Lys Val Phe Asn Thr Thr Pro Asp Asp Leu Asp Leu 335 340 345 His Val Ile Tyr Asp Val Ser His Asn Ile Ala Lys Val Glu Gln 350 355 360 His Val Val Asp Gly Lys Glu Arg Thr Leu Leu Val His Arg Lys 365 370 375 Gly Ser Thr Arg Ala Phe Pro Pro His His Pro Leu Ile Ala Val 380 385 390 Asp Tyr Gln Leu Thr Gly Gln Pro Val Leu Ile Gly Gly Thr Met 395 400 405 Gly Thr Cys Ser Tyr Val Leu Thr Gly Thr Glu Gln Gly Met Thr 410 415 420 Glu Thr Phe Gly Thr Thr Cys His Gly Ala Gly Arg Ala Leu Ser 425 430 435 Arg Ala Lys Ser Arg Arg Asn Leu Asp Phe Gln Asp Val Leu Asp 440 445 450 Lys Leu Ala Asp Met Gly Ile Ala Ile Arg Val Ala Ser Pro Lys 455 460 465 Leu Val Met Glu Glu Ala Pro Glu Ser Tyr Lys Asn Val Thr Asp 470 475 480 Val Val Asn Thr Cys His Asp Ala Gly Ile Ser Lys Lys Ala Ile 485 490 495 Lys Leu Arg Pro Ile Ala Val Ile Lys Gly 500 505 <210> SEQ ID NO 21 <211> LENGTH: 1929 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1841446 <400> SEQUENCE: 21 cggaagcaaa ggagccaaga ccatggcgaa agccggggat aagagcagca gcagcgggaa 60 gaaaagtcta aaacggaaag ccgctgccga agaacttcag gaggctgcag gcgctgggga 120 tggggcgacg gaaaacgggg tccaaccccc gaaagcggct gcctttccgc caggctttag 180 catttcggag attaaaaaca aacagcggcg acacttaatg ttcacgcggt ggaaacagca 240 gcagcggaag gaaaagttgg cagctaagaa aaaacttaaa aaagaaagag aggctcttgg 300 cgataaggct ccaccaaagc ctgtacccaa gaccattgac aaccagcgag tgtatgatga 360 aaccacagta gaccctaatg atgaagaggt cgcttatgat gaagctacag atgaatttgc 420 ttcttacttc aacaaacaga cttctcccaa gattctcatc acaacatcag atagacctca 480 tgggagaaca gtacgactct gtgaacagct ctccacagtt ataccaaact cacatgttta 540 ttacagaaga ggactggctc tgaaaaaaat tattccacag tgcatcgcaa gagatttcac 600 agacctgatt gttattaatg aagatcgtaa aaccccaaat ggacttattt tgagtcactt 660 gccaaatggc ccaactgctc attttaaaat gagcagtgtt cgtcttcgta aagaaattaa 720 gagaagaggc aaggacccca cagaacacat acctgaaata attctgaata attttacaac 780 acggctgggt cattcaattg gacgtatgtt tgcatctctc tttcctcata atcctcaatt 840 tatcggaagg caggttgcca cattccacaa tcaacgggat tacatattct tcagatttca 900 cagatacata ttcaggagtg aaaagaaagt gggaattcag gaacttggac cacgttttac 960 cttaaaatta aggtctcttc agaaaggaac ctttgattct aaatatggag agtatgaatg 1020 ggtccataag ccccgggaaa tggatacaag tagaagaaaa ttccatttat aaagtactga 1080 gagaatgata ttggattttg ctgaacaggc ctatcttgaa ctttggtaaa ttatttttga 1140 cagaatactc ttttcaaaat ggcatttgct gatttcataa acctttcacg tctggacgaa 1200 ttaccaaatg ccatgaattg ccactgtgtg tttatgtaga aaatacaaat aaaagttatt 1260 ttgatggctt aggtttcctt aaacttagtt ctcttgtttt tgggtaactg tgaataatta 1320 agttggaatc aagattcaga ttaactttcc tatttgcata gaacacatga gaggaataaa 1380 atggttggta aatattggct aacccttgat ttttatacca gattaacctt ggattcccag 1440 tgtctggcac agttttaata gcttaaatgg aggccaggtt tctggatgtt ttaacattct 1500 cttaagcctt cagaagggta aaaaatttaa agcaaaatga tctaccaggg tttaaagcaa 1560 agttgcaaat tactgaagct aatctttgct tcctgatttt gaggtttttg gttttttgtg 1620 cccacgttgt ggggagctct tttttacctc attacatggt gctgtagtac tccattcagg 1680 cactgaaaca aagttaaccc tataagtaac tcatggatgg aaacccgtag aacttaacag 1740 cctcctcctg accttaaaag aataaaggtt cacagtttac ctttaattcc ctagcagtct 1800 tgccagatgt atggcataaa gtcatgtgag aagagtaggt ggaaaaaact gtacaaactt 1860 aaccccttca ggtgttcaga acagattaat ataccatgta tttaatacca ataataatgc 1920 aaaataaag 1929 <210> SEQ ID NO 22 <211> LENGTH: 2113 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1850310 <400> SEQUENCE: 22 ctgccacgcc actgtgtcat gaaggaaagt gaaagggaac gaggaagtag gaatgcccac 60 gctcgttgac tccgtgggtg aatacagcag ttaggacata cacaccatca cctttgaaag 120 tgcttgtttg ggggagggaa ggacatacgg gtaactagaa ctacccagcg agtcgtccag 180 aggagaggat caggtttgag tcaggaggct ccctgtactg gagtcgtccc actattcctc 240 aagaaatctt agaaccagct tgtgaggaaa aacatttttt aatgtaataa aaatatgcca 300 ttattctttg aaatgccaaa tgatataaat attttgccta atacatattt attgtagatg 360 aaatgcactc ttctcgatga ggcctcgatt tgaatcaatg gggtgggcca caggaaatgt 420 cagaggaacc agaactcaga actcttcctc ctggaccttt cttcccttcc cttggaggta 480 tccctttgaa tcaggcctct ctcttctcat cagtctgtag cttcccccct tgtataacct 540 gctttccttt ttacatttat taaaagtgga ttttgtaaaa gcatttcatt gacacgcgac 600 ctatcacaga caatggaatt cgtcagtggt ggtaagactg aaatcctgat gcttttcaca 660 cttcttgtct cttgctatgt atttctgcct ctagccttgc catgttttgc cttttttttt 720 tctttttggc caattccttt ttatatgtgc ccacaacaga ggtggggaga cacggagcac 780 cctgggtcct tcccagcgct gctgggcagg ccccgtctcc aggccccagc tgttgaaact 840 ttgaagggca acaaacaacc atccacactg ccggacccta ggctgttcag ggaggcagct 900 catttccacc ccggccccag gacacccagc ctgtgcccca caaggatctc tctaaatggg 960 agggattgag gctacttttc tgccaagccc tattaagtag taatgtgggg aaacccactg 1020 tgtcagtgca ggaagcccta gacaaatgtt ttcaaataaa tttcactgcc cagcctgcac 1080 agatttccat ttgaagtact tcccatccac cctgacaccc aaaggggttt ttttgttttg 1140 ttttgttttt gagacagggt cttgctttgt tgcccaggct ggagtgcagt gacgtggtca 1200 tagctcactg cagcctcaac ctcctgggct caagtgaccc tcctgcctca gcctcccaaa 1260 gttctgagat gataggcatg agccattgtg cctagcctat tttgattttt ttcttagagt 1320 caaggtcttg ctctgttgcc caggctgatc ttggacttgc gagccaccat gcctggctgg 1380 gtttttttaa aaatagaatc tcactgatag cctgcaagaa acagatgcag tgcctgcttc 1440 cgtatcagtc caaggagccc tcgtgtttgc cacctttacc tttgaacctc cccctgcctc 1500 cctgcctgtg tccgcttttg cagctcaatg cagccatgac aaggaaagaa aagacaaagg 1560 aaggccagag agccgcgcag ttctctgcag gtgcagatgc aggcagtgga ggtggcctga 1620 gcaggcagaa ggacaccaag cgccctatgt tgcttgtcat tcatgacgtg gtcttggagc 1680 ttctgactag ttcagactgc cacgccaacc ccagaaaata ccccacatgc cagaaaagtg 1740 aagtcctagg tgtttccatc tatgtttcaa tctgtccatc taccaggcct cgcgataaaa 1800 acaaaacaaa aaaacgctgc caggttttag aagcagttct ggtctcaaaa ccatcaggat 1860 cctgccacca gggttctttt gaaatagtac cacatgtaaa agggaatttg gctttcactt 1920 catctaatca ctgaattgtc aggctttgat tgataattgt agaaataagt agccttctgt 1980 tgtgggaata agttataatc agtattcatc tctttgtttt ttgtcactct tttctctcta 2040 attgtgtcat ttgtactgtt tgaaaaatat ttcttctata aaattaaact aacctgcctt 2100 aagaaaaaaa aaa 2113 <210> SEQ ID NO 23 <211> LENGTH: 1652 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1887020 <400> SEQUENCE: 23 gacgaggtgc tattcgagtg ctagtggtgg aacgggacca cacgtattca caggcctcca 60 ctgggctctc agtaggtggg atttgtcagc agttctcatt gcctgagaac atccagctct 120 ccctcttttc agccagcttt ctacggaaca tcaatgagta cctggccgta gtcgatgctc 180 ctcccctgga cctccggttc aacccctcgg gctacctctt gctggcttca gaaaaggatg 240 ctgcagccat ggagagcaac gtgaaagtgc agaggcagga gggagccaaa gtttctctga 300 tgtctcctga tcagcttcgg aacaagtttc cctggataaa cacagaggga gtggctttgg 360 cgtcttatgg gatggaggac gaaggttggt ttgacccctg gtgtctgctc caggggcttc 420 ggcgaaaggt ccagtccttg ggagtccttt tctgccaggg agaggtgaca cgttttgtct 480 cttcatctca acgcatgttg accacagatg acaaagcggt ggtcttgaaa aggatccatg 540 aagtccatgt gaagatggac cgcagcctgg agtaccagcc tgtggaatgc gccattgtga 600 tcaacgcagc cggagcctgg tctgcgcaaa tcgcagcact ggctggtgtt ggagaggggc 660 cgcctggcac cctgcagggc accaagctac ctgtggagcc gaggaaaagg tatgtgtatg 720 tgtggcactg cccccaggga ccaggcctag agactccgct tgttgcagac accagtggag 780 cctattttcg ccgggaagga ttaggtagca actacctagg tggtcgtagc cccactgagc 840 aggaagaacc ggacccggcg aacctggaag tggaccatga tttcttccag gacaaggtgt 900 ggccccattt ggccctgagg gtcccagctt ttgagactct gaaggttcag agcgcctggg 960 ccggctatta cgactacaac acctttgacc agaatggcgt ggtgggcccc cacccgctag 1020 ttgtcaacat gtactttgct actggcttca gtggtcacgg gctccagcag gcccctggca 1080 ttgggcgagc tgtagcagag atggtactga agggcaggtt ccagaccatc gacctgagcc 1140 ccttcctctt tacccgcttt tacttgggag agaagatcca ggagaacaac atcatctgag 1200 catgtgtgct ctgcactggc tccactggct tgcatcctgg ctgtgttcac agccttgttt 1260 gctgcttcca tcttccccag tactgtgcca ggccttctcc ccctccccag tgtcctctcc 1320 tctcaggcag gccattgcac ccatatggct gggcaggcac aggcagtgag gccgaggcca 1380 atagcgagtg atgagcggga tcctaggact gatctgtagc ccatgctgat gtcacccacc 1440 agggcaatcc atctggaggc ctgagcaccc tggcccagga ctggcttcat cctggcactg 1500 accaggaaag actgcctctg accctcttag cagacagagc ccaggcatgg gagcactctg 1560 gggcagcctg gctcaggttt attgattttc gtctgtttac cctatccatt aatcaataca 1620 tgtaattaac tccttccaaa aaaaaaaaaa aa 1652 <210> SEQ ID NO 24 <211> LENGTH: 1120 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1911421 <400> SEQUENCE: 24 agagcgccgg aagcggtccg agaatgaaga gtgtgatcta ccatgcattg tctcagaaag 60 aggcgaatga ctccgatgtc cagccttcag gagcacagcg ggccgaggcc ttcgtgaggg 120 ccttcctgaa gcgcagcacg ccccgcatga gcccgcaggc ccgcgaggac cagctgcagc 180 gcaaggcggt ggtcctggag tacttcaccc gccacaagcg caaggagaag aagaagaaag 240 ccaaaggcct ctctgccagg caaaggaggg agctgcggct ctttgacatt aaaccagagc 300 agcagagata cagccttttc ctccctctcc atgaactctg gaaacagtac atcagggacc 360 tgtgcagtgg gctcaagcca gacacgcagc cacagatgat tcaggccaag ctcttaaagg 420 cagatcttca cggggctatt atttcagtga caaaatccaa atgcccctct tatgtgggta 480 ttacaggaat ccttctacag gaaacaaagc acattttcaa aattatcacc aaagaagacc 540 gcctgaaagt tatccccaag ctaaactgcg tgttcactgt ggaaaccgat ggctttattt 600 cctacattta cgggagcaaa ttccagcttc ggtcaagtga acggtctgcg aagaagttca 660 aagcgaaggg aacgattgac ctgtgaattc tttgccgtct aaggcagttg tttatgacag 720 ctgaaaactg gacactccct aaatgtccac ctttcagtga agagatagtt aagccaattc 780 catttataga ccacctccag ccagtgacgc tccgagttga ggatgttgaa caacatggga 840 aggtcgcagc gtactaagtg aagaagtcag aggacagagg aatttctctt tctaggagat 900 tttcattttg tgtgactccc atggggagga acagactggc aggaagcaca ccggggttaa 960 cactggttga cttgaatagg attattcgat ttttaaaaat acttttccat gttttctgag 1020 tgctctatga taaatcagtt gcatctgtga taatacagta catatgtgga cataaacagg 1080 gatcaaataa aggaggtatt gctgcaaaaa aaaaaaaaaa 1120 <210> SEQ ID NO 25 <211> LENGTH: 1000 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1911910 <400> SEQUENCE: 25 agtcctggaa agcgttgttg gcccggttgc tctggagccg ggtctcgggt ctggtggctg 60 ccggccctgc ggcatctcgc catggggagc acggagagca gcgagggccg cagggtgtcc 120 ttcggagtgg acgaggagga gcgggtccgg gtgctgcagg gtgtccggct gtctgaaaac 180 gtggtgaacc gcatgaagga gcccagctct ccaccccctg ctcccacatc ttctaccttt 240 ggccttcaag atggcaactt gagagcccct cacaaagaat ccacactgcc caggtcgggg 300 agcagtggtg gccagcagcc ctcagggatg aaggagggtg tcaagaggta tgaacaggag 360 catgctgcta tccaggataa gctcttccag gtggcaaaga gggaaagaga ggctgccacc 420 aagcactcca aggcatccct gcccacgggc gaaggcagca tcagccatga ggagcagaag 480 tcagtccggc tggccaggga gctggagagc agagaggcag agctaagacg ccgtgacacc 540 ttctacaagg agcagctgga gcgtattgag aggaagaatg ctgagatgta taaactgtct 600 tcagagcaat tccatgaggc agcctcaaag atggagagca caataaagcc ccgcagggtg 660 gagcccgtct gctcagggtt gcaggcccag attctccact gctaccgaga tcgcccgcat 720 gaggtgctgc tgtgctcgga cctggtcaag gcataccagc gctgcgtgag cgccgcccac 780 aagggctgag gagcagacat cattccctgc cctggcagtg acttggagcc ctgaagaagg 840 gaccaatcat gggaccacag ccactgtgcc ctgccgtttc ctgctgggcc cctgcatatg 900 cccctgagcc tggggctgcc acgtgtttag gaaacaaagt atgcgctact gtctgaaaac 960 aaataaagca gatgcctttg ttttcaaaaa aaaaaaaaaa 1000 <210> SEQ ID NO 26 <211> LENGTH: 2273 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 1928920 <400> SEQUENCE: 26 gggggtgaag cgatacgttt tgcccgcatt cggggcgcgc ggactggggg ggtccctgtg 60 gggctcccgg agttaagatg gcgtcctcag cggaggggga cgaggggact gtggtggcgc 120 tggcgggggt tctgcagtcg ggtttccagg agctgagcct taacaagttg gcgacgtccc 180 tgggcgcgtc agaacaggcg ctgcggctga tcatctccat cttcctgggt tacccctttg 240 ctttgtttta tcggcattac cttttctaca aggagaccta cctcatccac ctcttccata 300 cctttacagg cctctcaatt gcttatttta actttggaaa ccagctctac cactccctgc 360 tgtgtattgt gcttcagttc ctcatccttc gactaatggg ccgcaccatc actgccgtcc 420 tcactacctt ttgcttccag atggcctacc ttctggctgg atactattac actgccaccg 480 gcaactacga tatcaagtgg acaatgccac attgtgttct gactttgaag ctgattggtt 540 tggctgttga ctactttgac ggagggaaag atcagaattc cttgtcctct gagcaacaga 600 aatatgccat acgtggtgtt ccttccctgc tggaagttgc tggtttctcc tacttctatg 660 gggccttctt ggtagggccc cagttctcaa tgaatcacta catgaagctg gtgcagggag 720 agctgattga cataccagga aagataccaa acagcatcat tcctgctctc aagcgcctga 780 gtctgggcct tttctaccta gtgggctaca cactgctcag cccccacatc acagaagact 840 atctcctcac tgaagactat gacaaccacc ccttctggtt ccgctgcatg tacatgctga 900 tctggggcaa gtttgtgctg tacaaatatg tcacctgttg gctggtcaca gaaggagtat 960 gcattttgac gggcctgggc ttcaatggct ttgaagaaaa gggcaaggca aagtgggatg 1020 cctgtgccaa catgaaggtg tggctctttg aaacaaaccc ccgcttcact ggcaccattg 1080 cctcattcaa catcaacacc aacgcctggg tggcccgcta catcttcaaa cgactcaagt 1140 tccttggaaa taaagaactc tctcagggtc tctcgttgct attcctggcc ctctggcacg 1200 gcctgcactc aggatacctg gtctgcttcc agatggaatt cctcattgtt attgtggaaa 1260 gacaggctgc caggctcatt caagagagcc ccaccctgag caagctggcc gccattactg 1320 tcctccagcc cttctactat ttggtgcaac agaccatcca ctggctcttc atgggttact 1380 ccatgactgc cttctgcctc ttcacgtggg acaaatggct taaggtgtat aaatccatct 1440 atttccttgg ccacatcttc ttcctgagcc tactattcat attgccttat attcacaaag 1500 caatggtgcc aaggaaagag aagttaaaga agatggaata atccatttcc ctggtggcct 1560 gtgcgggact ggtgcagaaa ctactcgtct cccttttcac agcactcctt tgccccagag 1620 cagagaatgg aaaagccagg gaggtggaag atcgatgctt ccagctgtgc ctctgctgcc 1680 agccaagtct tcatttgggg ccaaagggga aacttttttt tggagaaggc gtcttgcttt 1740 gtcacccacg ctggaatgca gtggcgggat ctcagctcac cgcaacctcc acctcctggg 1800 ttcaagtgat tttcctgcct cagcctccca agtagctggg aatacaggca cgccaccatg 1860 cccagctaat ttttgtattt tcagtagaaa cgggatttca ccacgttggc caggctggtc 1920 tcgaactcct gaccgcaagt gatccacccg cctccgcctc ccaaagtgct gggattacag 1980 gcgtgagcca ccgtgcccgg cccaaagggg aaactcttgt gggaggagca gaggggctca 2040 catctcccct ctgattcccc catgcacatt gccttatctc tccccatcta gccaggaatc 2100 tattgtgttt ttcttctgcc aatttactat gattgtgtat gtgccgctac caccaccccc 2160 cccatggggg ggtggagagg ggtgcaaggc cctgcctgct ccactttttc taccttggaa 2220 ctgtattaga taaaatcact tctgtttgtt cagtttttca aaaaaaaaaa aaa 2273 <210> SEQ ID NO 27 <211> LENGTH: 925 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2170846 <400> SEQUENCE: 27 cgcagaaaca gcctatagac gccacgagtc ggcggcgcta ccgaggggct gtgggcgcgc 60 agctggaacc tccggctgtc agtgcgctta cagttcctaa ccccgaccct gcgcgcaccc 120 gcactatggc agccccgccg cagctaaggg ctctgctcgt agtcgtcaac gcactgctgc 180 gcaagcgccg ctaccacgct gcgttggccg tgcttaaggg cttccggaac ggggctgtct 240 atggagccaa aatccgggcc cctcacgcgc tggtcatgac ctttctcttc cggaatggca 300 gcctccagga gaagctgtgg gccatactgc aggccacata tatccactcc tggaacctgg 360 cacggtttgt gttcacctac aagggtctcc gtgccctgca gtcctacata caaggcaaga 420 cctacccagc acacgcattc ctggcggcct tcctcggggg tatcctggtg tttggagaaa 480 acaataacat caacagccag atcaacatgt acctgttgtc acgcgtcctg tttgccctga 540 gccgcctggc tgtagagaag ggctacatcc ctgaacccag gtgggacccg ttcccgctgc 600 tcactgcggt ggtgtggggg ctggtgctgt ggctctttga gtatcaccga tccaccctgc 660 agccctcgct gcagtcctcc atgacctacc tctatgagga cagcaatgta tggcacgaca 720 tctcagactt cctcatctat aacaagagcc gtccctccaa ttaatgcagc cctgaggtgt 780 ctggctgtgg ctcaagattt ggccccatgc agaccctccc aaaggatact gccttctcaa 840 gatcataggc ctcagactcc aactggtgtt atcccagggt tctgtttgct gaagtaaaaa 900 cactgatttt aaaaaaaaaa aaaaa 925 <210> SEQ ID NO 28 <211> LENGTH: 1570 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2176361 <400> SEQUENCE: 28 gggcctccgt ctccgacgct gacttcctca gcgcaccagc actgtctccg gtggcaagtc 60 gaccagctcc tcccacacgg attctgtgta gaaatccacg gtatgtgcat tggaaatgga 120 cagggcatcc ctcaggaact gcagcagtcc ctgcaacttg gcacgcagcg tgggcaggtc 180 cggggtcacc gggagagggc aagaagccgc catgacgctc accctccgag gctgtaggtg 240 gcgcaaacat ggccgtgagg cgccagctcg gcccaaatta gcacaggcag gctcagtaga 300 gctccgagcc ggattccttc tgagcgattg attgcttcgg cttggtgacg tattttgtgg 360 gcgtccgtcg tggtcttctg atgacgcatt agtcggctgc aatggcgccg gtgaggcggt 420 ccgcgaagtg gcggcctggt ggtattgagg cgcgtggtga aggggtttcc actgtcgggt 480 acaggaataa gaatgtgaga cagaagacat ggcggcctaa ccacccgcaa gccttcgtgg 540 ggagcgttcg cgagggacaa ggctttgctt ttcgaagaaa actgaaaata cagcaaagtt 600 acaagaaatt gctacggaag gaaaagaagg ctcaaacgtc actggaatct caattcacag 660 atcgataccc agataatctg aaacatctct atttagctga agaggaaaga cataggaagc 720 aagcaagaaa agtcgaccat cctttgtcag aacaagttca ccagccgttg cttgaagaac 780 agtgtagcat tgacgagcct ttatttgaag atcagtgtag ctttgaccag cctcagccag 840 aagaacaatg tattaaaaca gtaaactcct ttacaattcc aaagaaaaat aaaaagaaaa 900 catcaaatca aaaagcacaa gaagaatatg aacagataca agccaaacgt gctgctaaga 960 aacaagaatt cgagaggaga aaacaggaga gagaagaagc ccaaaggcag tacaaaaaga 1020 agaaaatgga agtgtttaaa atactgaaca aaaagactaa aaagggccaa ccaaacttga 1080 atgtacaaat ggagtacctt cttcaaaaaa tacaagaaaa atgttaaaca ttttgttcct 1140 acaggttaaa atatctgctg cctattaggt tcttctgtga catgtgcctc ccagcagtga 1200 actaaatttg tcgacataaa ctggattgct aaactatgct aaatataaga tgttcacata 1260 tttttattat ggtaaaaaat tttctaaata tgttctacat gtttcttatt tatttgcctc 1320 tgaaggaagg ttggcctgaa gaactgaaag aacctcttat tttgcaagac aggcccaagc 1380 atgtaatact tttgtaccat atgagattta tatgaaataa attttttaaa aataaggaat 1440 cagagctatc aatgaagcat ttcaatgaaa tatttcaatt tagtaaacag ttacgttgtt 1500 ttaaaattta ttttaatgat gagggaggca aagactcctc ttggactttt tatttatttt 1560 aagtacatgt 1570 <210> SEQ ID NO 29 <211> LENGTH: 1868 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2212732 <400> SEQUENCE: 29 tcaccatgag gatccagccc ttggtaaggc cgggactgca gaagcagtca tccctgaaaa 60 ccatgaggtt ctggcaggcc cagatgagca ccctcaggac acagatgcaa gagatgctga 120 tggggaggct agagatcggg agccagcaga ccaagctttg ctgcttagcc agtgtgggga 180 taaccttgag tcccctctgc ctgaagctag ctcagctcca ccggggccaa cccttgggac 240 actgcctgaa gtagagacaa taagggcatg ctccatgccc caggagcttc ctcagtcccc 300 caggacccga cagcctgagc cagatttcta ctgtgtcaag tggatccctt ggaaaggaga 360 acagacaccc atcatcaccc agagcactaa cggcccttgc cctctccttg ccatcatgaa 420 catcctcttt cttcagtgga aggtgaagct ccccccgcag aaggaagtga tcacatcgga 480 tgagctcatg gcccatcttg gaaactgcct cctgtccatc aagccccagg agaagtcaga 540 gggacttcag cttaattttc agcagaatgt ggatgatgca atgacagtgc tgcctaaact 600 ggccacaggt ctggatgtca atgtgcgatt cacaggcgtc tctgattttg agtatacacc 660 cgagtgcagt gtctttgacc tgctaggcat acctctgtac catggctggc ttgttgatcc 720 acagcagagt cctgaggctg tgcgtgcagt tgggaaactg agttacaacc agctggtgga 780 gaggatcatc acctgcaaac actccagtga caccaacctc gtgacagaag gcctgattgc 840 agagcagttc ctggagacca ccgcggccca gctgacctac cacggactgt gtgagctgac 900 agcagctgct aaggagggtg aacttagcgt ctttttccga aacaaccact ttagcaccat 960 gactaagcat aagagtcact tatacctact ggtcactgac cagggctttc tacaggagga 1020 gcaagtcgta tgggagagcc tgcacaatgt ggatggagac agctgctttt gtgactctga 1080 ctttcacctg agtcattccc tgggcaaggg gcctggagca gaaggtggga gtggctcccc 1140 agaaaagcag ctgcaggtag accaggacta cctgattgct ctgtccctgc agcagcaaca 1200 gccacgaggc ccgctggggc ttaccgactt ggagctggcc cagcagcttc agcaagagga 1260 gtatcaacag cagcaggcag cgcagccagt gcggatgcgg acgcgggtcc tgtcactgca 1320 ggggagagga gccacatctg gacgcccagc cggggagcgt cggcagaggc cgaagcacga 1380 gtcagactgc attctgctgt agctctgccc cagtgccagg ctggcctgcc ccttcttcca 1440 gaggctatgg ctagttggct tgctcccccg cctccacccc tgagatgtgc tggataactt 1500 atttatggac tgttggggat gagagcaggc aacaaatgcc aaggtcagac ttggtaatgt 1560 ccttgacctc acgtgctgct gccttctctg cctcccaccc agggcaacac taggattggt 1620 gggtttctgg ttctcaactc ccggtccctg aatagtcaca cgtatgtaca gactgaggct 1680 ctggggtgag gtccctatcc agaatgcatc tcttctgctt cccatccctg ctgcctggat 1740 gctcctgatc acctaggcag gcctgtctcc agttgtttca gagcttaatt tgggtttcta 1800 tctcttattt gtaatgcctt cctggggttt ggaaataaaa cttctggccg ggcaaaaaaa 1860 aaaaaaaa 1868 <210> SEQ ID NO 30 <211> LENGTH: 1401 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2303457 <400> SEQUENCE: 30 ccaagctgca gctggcaggg attgcggggt gccggccgtc tgagtttttt taaaactgct 60 cgccgcgaag tctgtctgca gccaaaatgt ccaacagaaa caacaacaag cttcccagca 120 acctgccgca gttacagaat ctaatcaagc gagacccgcc ggcctacatc gaggagtttc 180 tacagcagta taatcactac aaatccaatg tggagatttt caaattgcaa ccaaataaac 240 ccagcaaaga actagcagag ctggtgatgt ttatggcaca gattagtcac tgctacccag 300 agtacctaag taattttcct caagaggtga aagatcttct ctcctgcaat cataccgtat 360 tggatccaga tctgcgaatg acattttgca aagctttgat cttgctgaga aataagaatc 420 tcatcaatcc atcaagcctg ctagaactct tctttgaact ttttcgttgc catgataaac 480 ttctgcgaaa gactttatac acacatattg tgactgatat caagaatata aatgcaaaac 540 acaagaacaa taaagtgaat gtagtattgc aaaatttcat gtacaccatg ttaagagata 600 gcaatgcaac cgcagccaag atgtctttag atgtaatgat tgaactctac agaaggaaca 660 tctggaatga tgcaaaaact gtcaatgtta tcacaactgc atgtttctct aaggtcacca 720 agatattagt tgccgctttg acattctttc ttgggaaaga tgaagatgaa aaacaggaca 780 gtgactccga atctgaggat gatggaccaa cagcaagaga cctgctagta caatatgcta 840 cagggaagaa aagttccaaa aacaagaaaa agttggaaaa ggcaatgaaa gtgctcaaga 900 aacaaaaaaa gaagaaaaaa ccagaggtgt ttaacttttc agccattcac ttgattcatg 960 atccccaaga ttttgcggaa aaactactaa agcagcttga gtgctgtaag gagaggtttg 1020 aagtgaagat gatgctcatg aaccttatct ccagattggt gggaattcat gagcttttcc 1080 tcttcaattt ctatcccttt ttgaaaaggt ttctgaagcc ccaccaaagg gaggtaacca 1140 agatccttct gtttgttgaa aaagattctc atcacttagt accccaaggg ttttttaatt 1200 catggttaat gcttggggaa aaaatttttt ttaacggaaa aaaatctggg aaaatgttaa 1260 tgacagttgg gaatttaatg gttaaaagag gggtatataa acgctccaaa gtgttccttg 1320 ggggaaatag tgttggaaga aatttttttc aaaaaaatcc agggggctca agttaaaaaa 1380 accccccagg cgggtgtgta t 1401 <210> SEQ ID NO 31 <211> LENGTH: 1409 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2317552 <400> SEQUENCE: 31 gtcggggctg cgccgtacaa cttccggctg taaagatggc ggcttcctag tgagtcggcg 60 gctgatttag aaggaggttc aggctacggt gagccgaagc cacacaggag ccatggaagt 120 ggcagagccc agcagcccca ctgaagagga ggaggaggaa gaggagcact cggcagagcc 180 tcggccccgc actcgctcca atcctgaagg ggctgaggac cgggcagtag gggcacaggc 240 cagcgtgggc agccgcagcg agggtgaggg tgaggccgcc agtgctgatg atgggagcct 300 caacacttca ggagccggcc ctaagtcctg gcaggtgccc ccgccagccc ctgaggtcca 360 aattcggaca ccaagggtca actgtccaga gaaagtgatt atctgcctgg acctgtcaga 420 ggaaatgtca ctgccaaagc tggagtcgtt caacggctcc aaaaccaacg ccctcaatgt 480 ctcccagaag atgattgaga tgttcgtgcg gacaaaacac aagatcgaca aaagccacga 540 gtttgcactg gtggtggtga acgatgacac ggcctggctg tctggcctga cctccgaccc 600 ccgcgagctc tgtagctgcc tctatgatct ggagacggcc tcctgttcca ccttcaatct 660 ggaaggactt ttcagcctca tccagcagaa aactgagctt ccggtcacag agaacgtgca 720 gacgattccc ccgccatatg tggtccgcac catccttgtc tacagccgtc caccttgcca 780 gccccagttc tccttgacgg agcccatgaa gaaaatgttc cagtgcccat atttcttctt 840 tgacgttgtt tacatccaca atggcactga ggagaaggag gaggagatga gttggaagga 900 tatgtttgcc ttcatgggca gcctggatac caagggtacc agctacaaat atgaggtggc 960 actggctggg ccagccctgg agttgcacaa ctgcatggcg aaactgttgg cccaccccct 1020 gcagcggcct tgccagagcc atgcttccta cagcctgctg gaggaggagg atgaagccat 1080 tgaggttgag gccactgtct gaaccatccc tgtacatctg caccttcttg tgcaaggaag 1140 tccttggcct aaagccttgg ttctcaaact gggttccttg ggacctccgg ggtggggggg 1200 ttccaggagg cacgtagggt accttgcagg gtcctaggag ggaaacccag gattccagga 1260 gggatcccag gaactgtggg cacccatttt ctgtgtctcc cagcccattt ccactcctag 1320 tttgtcatgg ataatttttg ttcttccctg tgtgattttt gccatcaaaa taaaaatttg 1380 agactcgtta accgaaaaaa aaaaaaaaa 1409 <210> SEQ ID NO 32 <211> LENGTH: 1888 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2416366 <400> SEQUENCE: 32 tgtgattcgg agagtgagga tccctcaagg aaccaggcta gtgattcggg aaatgaggag 60 ctactcaaac cccgagtcag tgactctgag agtgaggggc ctcagaaggg gcctgccagt 120 gactcagaaa ctgaggatgc gtccagacac aaacagaagc cagagtcaga tgatgacagc 180 gacagggaga ataagggaga ggatacagaa atgcagaatg actccttcca ttcagacagc 240 catatggaca gaaaaaagtt tcacagttct gatagtgagg aggaagaaca caaaaagcaa 300 aaaatggaca gtgatgaaga tgaaaaagag ggtgaggagg agaaagtagc gaagagaaaa 360 gctgctgtgc tttctgatag tgaagatgaa gagaaagcat cagcaaagaa gagtcgtgtt 420 gtctctgatg cagatgactc tgacagtgat gctgtatcag acaagtcagg caaaagagag 480 aagaccatag catctgacag tgaggaagaa gctgggaaag aattgtctga taagaaaaat 540 gaagagaagg atctgtttgg gagtgacagt gagtcaggca atgaagaaga aaatcttatt 600 gcagacatat ttggagaatc tggtgatgaa gaggaagaag aatttacagg ttttaaccaa 660 gaagatctgg aagaagaaaa aggtgaaaca caggtaaaag aagcagaaga ttcagattct 720 gatgataaca taaagagagg aaaacatatg gactttctgt cagattttga gatgatgttg 780 cagcgaaaaa agagcatgag tggcaagcgc agacggaacc gcgatggtgg cacctttatt 840 agtgatgcag acgacgtcgt gagtgccatg atcgtcaaga tgaatgaagc tgctgaggaa 900 gacagacagt tgaacaatca aaaaaagcca gcactgaaaa aattaacttt actgcctgct 960 gtagttatgc accttaagaa gcaggacctt aaagaaacat tcattgacag tggtgtgatg 1020 tctgccatca aagaatggct ctcacctcta ccagatagga gtttgcctgc actcaagatc 1080 cgggaggagc tgctgaagat cctgcaagag ctgcctagtg tgagccagga gaccctgaag 1140 catagtggga ttggacgagc agtgatgtat ctctataaac accccaagga gtcaaggtct 1200 aacaaggaca tggcagggaa attaatcaat gagtggtcta ggcctatatt tggtcttacc 1260 tcaaactaca aaggaatgac aagagaagaa agggagcaga gagatctaga acagatgcct 1320 caacgacgaa gaatgaacag cactggtggt cagacaccca gaagagacct ggaaaaggtg 1380 ctgacaggag aggagaaggc tcttagacct ggagatcctg gattctgtgc ccgtgcaagg 1440 gtcccaatgc cttcaaacaa ggactatgtt gtcaggccca aatggaatgt ggaaatggag 1500 tcatccaggt ttcaggcgac ctccaagaag ggtatcagtc gactggataa acagatgaga 1560 aagttcacag atataaggaa aaaaagcaga tctgcacacg cagtgaaaat cagcattgag 1620 ggcaacaaaa tgccattgtg accttgcctg gaatgtgtcc ccatctctac tctaagaaat 1680 gcgcaatgga ctctttggag aaagaagata ttttaaaaca tttttagtgt gtctgtaaat 1740 ggttcagcgt gtatcagatg ttgtcatagg actcacattt ctctcagtta tatttaaaac 1800 cgttgtgtac tttgtacaaa ggaatactag tcatacttct ataaacttta cacaataaaa 1860 tttcattctg gttaaaaaaa aaaaaaaa 1888 <210> SEQ ID NO 33 <211> LENGTH: 1897 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <221> NAME/KEY: unsure <222> LOCATION: 1892 <223> OTHER INFORMATION: a or g or c or t, unknown, or other <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2472980 <400> SEQUENCE: 33 ttattgaaga catggccgcc gcgtattttc cagactgcat agtcagaccc tttggctcct 60 cagtcaacac ttttgggaag ttaggatgtg atttggacat gtttttggat ctagatgaaa 120 ccagaaacct cagcgctcac aagatctcag gaaattttct gatggaattt caagtgaaaa 180 atgttccttc agaaagaatt gcaactcaga agatcctgtc tgtgttagga gagtgccttg 240 accactttgg ccctggctgt gtgggtgtgc aaaaaatatt aaatgcccgg tgtccgctcg 300 tgaggttctc acaccaggcc tccggatttc agtgtgattt gactacgaac aataggattg 360 ccttgacaag ttccgaactc ctttatatat atggtgccct agactcaaga gtgagagcct 420 tggtgttcag tgtacggtgc tgggctcgag cacattcact aacaagtagt attcctggtg 480 catggattac aaatttctcc cttacaatga tggtcatctt ttttctccag agaagatcac 540 cccctattct tccaacacta gattccttaa aaaccctagc agatgcagaa gataaatgtg 600 taatagaagg caacaactgc acatttgttc gtgacttgag tagaattaaa ccttcacaga 660 acacagaaac attagaatta ctactgaagg aattttttga gtattttggc aattttgctt 720 tcgataaaaa ttccataaat attcgacagg gaagggagca aaacaaacct gattcttctc 780 ctctgtacat tcagaatcca tttgaaactt ctctcaacat aagcaaaaat gtaagtcaaa 840 gccagctgca aaaatttgta gatttggccc gagaaagtgc ctggatttta caacaggaag 900 atacagatcg accttccata tcaagtaatc ggccctgggg gctggtatcc ctattgctac 960 catctgctcc aaacagaaag tcctttacca agaagaaaag caataagttt gcaattgaaa 1020 cagtcaaaaa cttgctagaa tctttaaaag gtaacagaac agaaaatttc acaaaaacca 1080 gtgggaagag aacaattagt actcagacat gatggctgct acattgtgta aagaactggg 1140 cttagcctat caaatggtct gtggacttac ttggaaaaac tgatttgaaa ctttcacaga 1200 tctcagcttt catctgatgt cacttttcat gatcttctca ttggccccct taacctggtc 1260 tgaagttctg ggatgttttc agtttgatca gtctgatact cagtggcact ttattaaaac 1320 atcagctgtg gagtgtggcg gtgcacacct gtagtcccag ctgctcagga ggctgaggca 1380 ggaggatctc ttgagcccag gattttgaat ccatcgtgga caacatagca agattccatc 1440 tctaaaaaaa atgaaaataa acataagcca caaggaatgg gtgaaagatt attgtaatgt 1500 gctttaacta aataggtaaa tatactaaac aaatgctaaa actcagtttt aggatgaaac 1560 cattgttgat atccacatca gtccctgttt agaaaacatt taaaatgact tttagttatg 1620 tacagtacgt tggcaatgaa tacattaagc ttcaaaattt ggtagtgctc tcgaatatgt 1680 atatttgtat ttttcaagcg aagttctctt attcacatat aaattaaagt gggttggtac 1740 tgatatcaaa aaatgtttat gtttttagaa cagacatttc agtcactgca ttcttaggta 1800 ttccaaacca aatatgatga catcaataga ttgcatttta aaaatattgt ttgatttttc 1860 tatggtcaaa aataaaattg tgttctactt tnctggt 1897 <210> SEQ ID NO 34 <211> LENGTH: 1132 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2541640 <400> SEQUENCE: 34 gtggagcctc gaacccgaga cgctagaccc aatttggtgc ttatgggagg agtgggagtg 60 gcagaggcag cgagaccact cctttcttgg cccaccattt ctctcaccat ttttactgca 120 gtgaactcct ctcagggtgg ggggctggta cagaggcaac tccgctttca taactctcac 180 agggtccttt gtcggaggtg tccctgccct cctaccccag cgtggtggga gtgcgacgcc 240 aggcttctcc ctcctccctg gcctcctgtg cctccagcct ccacatctcc tgagatcctc 300 cccaccccac acttacacag gtccccacac gcccctggtg cccctaaacc gccacccaac 360 cccactcacc ctggggccgg gactggggtg tcagagctga gccagggacc ttgggaggtg 420 gctgggaccg gcgcctcctg ctccctcttt cattttccct ttcggatttg gccaggatgg 480 aggacaggac aggatggctg agaggggcta gcgctccccc ttctcttaaa ggaacagggt 540 gaccctgctg gctggctccg cccttgggaa caggcacgga ccacgcccct tgctgggctc 600 tgtggccctc actctttagc taaatttcca tctggaaatg ggggaggggg gtaaccagag 660 ccacatcctt tatcgaggaa gctgggggat tcctagaagc ccttcccaga cagaccctca 720 gtctggagcc tagatccact acgcactaac catgccctgt gaagatggtc ttcggggaca 780 gagggaggtt tggtgctgga gtttaagggg cagcaaacat ctaacagctt ctggttccag 840 ctctgccacc gctcaccttg gacaaatcat tcggcggtac tgagccaggc tttcctgcct 900 tgtcaacaaa ggcggccagt ccagctgacc cccaagtttc cttccagctt tacatctttt 960 ttggaacggg gtctctctac atagcccagg tcgatcttga actcctggct tcaagtgatc 1020 ctcacacctt agcctcctga gtagctggga ttctctactt cttaatattg aattcccaac 1080 catattacag agaaccaaaa aaacccttgt gcctgcaaat aaataaaaaa aa 1132 <210> SEQ ID NO 35 <211> LENGTH: 1763 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <221> NAME/KEY: unsure <222> LOCATION: 1612,1613, 1737, 1745 <223> OTHER INFORMATION: a or g or c or t, unknown, or other <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2695204 <400> SEQUENCE: 35 aggctacagc tagggctgct cagtaactgc cccaaccact accacgtccc caagtgcaac 60 gggacccacg gaactacagg gtcctgctga gctcaggagc tgcacggaat ggtggcagtc 120 accttgccag tgcagcaggc atggatcaga ctgcaagcta ggggcacgag gcatcagttg 180 ggaagagggg acgcacagct aggcttgagg cctcttcatc gggatgtccc caggcccccc 240 agcccaggcc cagcataaag gccgtgttgg ggggcccccc tgacccaagg ggggcttcat 300 gcgccacgtg caggcggagc gtagtccatc ctcagagccg gacgtggccc ttcacagcct 360 ccagtcaggc agggggccct ccagggtggc ctgctcatgg gctacagccc agcagggggg 420 gcgacatccc ccggggtcta ccaggcccct gccccaccaa gcgaaggctg cttcctgctg 480 gagaagcccc agatgtcagc tctgaggaag aggggccagc ccctcggagg cgccggggat 540 ccctgggcca ccctactgct gccaacagtt ctgatgccaa agccacaccc ttctggagcc 600 acctgctgcc tgggcccaaa gagcctgttt tggacccaac agactgcggt cccatggggc 660 ggaggctgaa aggagcccgt cgcctgaagc tgagccccct tcgaagcctc cggaaggggc 720 caggcctgct gagccccccc agtgcctccc ctgttcctac ccctgctgtc agccgtaccc 780 tgctgggcaa ctttgaggaa tcattgctgc gaggacgttt tgcaccatct ggccacattg 840 agggcttcac agcagaaatt ggagctagtg ggtcatactg cccccagcac gtcacgctgc 900 ctgtcactgt cacattcttt gatgtttctg agcaaaatgc cccggctccc ttcctgggca 960 tcgtggatct gaaccccttg gggaggaagg gttacagcgt gcccaaggtg ggcaccgtcc 1020 aagtgacctt atttaacccc aaccagactg tggtaaagat gttccttgtg acctttgact 1080 tctcggacat gcctgctgcc cacatgacct tcctgcgcca tcgcctcttt ttggtgcctg 1140 tgggtgagga gggaaatgct aaccccaccc accgcctcct ctgctacttg ctgcacctca 1200 ggttccggag ctcccgctca ggccgcttaa gcctgcatgg agatatccgc ctgctttttt 1260 cccgccggag cctggagctg gacacagggc tcccctacga actgcaggct gtgaccgagg 1320 cccctcataa tccacgttat tcacctttgc cctgattgcc agcactctga acccatgcgg 1380 gctaatgacc tgcccatcct gctccatctt agagaacata tatggagaga cagcaagaga 1440 cccttcaggc ttgaattaaa gccctcacca tgctcacgcc caaatggatt atttgggtgt 1500 ttaaagcttc tgattcttac tacaccctgc cctacttcgg gtactccatg tgcctgtccc 1560 ctcccttggg tttcccagga cagcttaggt agtagggagg aactggagct annctgagat 1620 tttctcaagt tcccaggcaa gttatgccag cgttgcctcc ctgtcctggg caggggccac 1680 ttgttatttt atttattttt aatttataat ttattcaatt ggattgcctt ggtaaantcc 1740 cacanctgat aattggcatc act 1763 <210> SEQ ID NO 36 <211> LENGTH: 2110 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2805526 <400> SEQUENCE: 36 cagagaacgc gctcgcgtag aggaattccg ggtcctcctc ctcgccctac cttggtttac 60 ctgcagccgc ctagtcctct tctccttctc tttgatgatt cagggcctcg gcttccctca 120 cgatattgca gaaagacaca gctttcctct tcctctccaa accacctcga agcaggggca 180 taattggaat atcatttgga gaaagtgtca tggaagttct gcgtccacag cttataagaa 240 ttgatggccg gaattacagg aagaatccag tccaagaaca gacctatcaa catgaagaag 300 atgaagagga cttctatcaa ggctccatgg agtgtgctga tgagccctgt gatgcctacg 360 aggtggagca gaccccacaa ggattccggt ctactttgag ggcccccagc ttgctctata 420 agcatatagt tggaaagaga ggggacacta ggaagaaaat agaaatggag accaaaactt 480 ctattagcat tcctaaacct ggacaagacg gggaaattgt aatcactggc cagcatcgaa 540 atggtgtaat ttcagcccga acacggattg atgttctttt ggacactttt cgaagaaagc 600 agcccttcac tcacttcctt gcctttttcc tcaatgaagt tgaggttcag gaaggattcc 660 tgagattcca ggaggaagta ctggcgaagt gctccatgga tcatggggtt gacagcagca 720 ttttccagaa tcctaaaaag cttcatctaa ctattgggat gttggtgctt ttgagtgagg 780 aagagatcca gcagacatgt gagatgctac agcagtgtaa agaggaattc attaatgata 840 tttctggggg taaaccccta gaagtggaga tggcagggat agaatacatg aatgatgatc 900 ctggcatggt ggatgttctt tacgccaaag tccatatgaa agatggctcc aacaggctac 960 aagaattagt tgatcgagtg ctggaacgtt ttcaggcatc tggactaata gtgaaagagt 1020 ggaatagtgt gaaactgcat gctacagtta tgaatacact attcaggaaa gaccccaatg 1080 ctgaaggcag gtacaatctc tacacagcgg aaggcaaata tatcttcaag gaaagagaat 1140 catttgatgg ccgaaatatt ttaaagttgt ttgagaactt ctactttggc tccctaaagc 1200 tgaattcaat tcacatctct cagaggttca ccgtagacag ctttggaaac tacgcttcct 1260 gtggacaaat tgacttctcc tgaggtggat cttggaaagc actagaaact aaacatcttc 1320 accaggtgct gaagaaaagt gtcttcgttt taattgccaa gcagggatgt ggacatttgg 1380 atggtgactt tcctgggtgg ttccccatag attcaccatt gcctctaatg gtgtctacac 1440 ccgtcatact accagctgag atggtggtgg gcataaggag aatttgtgcc tataaccctt 1500 agtgtgttct ggtttttttt cttttaattt ttaaattgtc gtaaaatact cataaaacat 1560 actgtcttca ccatttttaa gtgcacagtt cagtaacgtt aactgttaat acattcataa 1620 tgctgtgtgg ccgtcaccgc cgtccatctc cataggcttc tcagcttgta aaatggaaac 1680 tgtacccatt aaacagtaat tcccactcct cccagccccc gcagccacca ttctgctttc 1740 tgtctctctg gttttgacta ttctcagtat ctcatataag tggaatcata cagtgacttg 1800 tctttttgtg actggcttat ttcacttagc ataatgtcct caaggttcat ccatgttgtg 1860 tcaggtgaca ggatttcctt cctgtattat acatataatg gaatgttccg ttacatgtgc 1920 gtaccacact ctgttcctcc atcagtgaac acttggcttg cttcctcttg actattggga 1980 gtagtgctaa tacagacacg ggggtgcaaa tatctctttg agactctgcc tttaattcct 2040 ttaccaccca gagttttatc tttagcaaaa taaccattaa agttgtttgc cttttaaaaa 2100 aaaaaaaaaa 2110 <210> SEQ ID NO 37 <211> LENGTH: 1493 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2850382 <400> SEQUENCE: 37 tggcggtggc gcggaccacg gcaggagcgg accgggcccg gggctgggcc ggcgtcgaac 60 gcagcgaagg ccggaggatg gaaccaggtg aggagctgga agaggagggc tctccaggtg 120 gccgtgagga tggcttcacc gccgagcacc tggctgcaga ggccatggca gctgacatgg 180 acccctggct agtgtttgat gcccgcacaa cgcctgccac tgagctggat gcctggctgg 240 ccaagtaccc accatcccaa gttacccgct atggggaccc cggttcaccc aactcagagc 300 ctgtgggctg gattgcagtg tatgggcagg gctacagccc caactccggg gacgtgcagg 360 gcctgcaggc agcctgggaa gctctgcaga ccagtgggcg gcccatcaca ccgggtaccc 420 tgcgccagct cgccatcacc caccacgtgc tctcgggcaa gtggcttatg catctggcac 480 cgggcttcaa gctggaccac gcctgggctg gcattgcccg ggccgtggtt gaaggccggc 540 ttcaggtggc caaggtgagc ccacgtgcca aggagggtgg gcgccaggtc atctgtgttt 600 acacggacga cttcacggac cgcttgggtg tactggaggc ggattcagcc atccgtgcag 660 cgggcattaa gtgcctgctc acctacaagc ctgatgtcta cacctacctg ggcatctacc 720 gggccaatcg ctggcacctc tgccccactc tctatgagag ccgtttccag cttgggggta 780 gtgcccgtgg ctcccgagtg cttgaccgtg ccaacaacgt ggaactgacc tagaggggcc 840 aaattgggga gactgcccac tcccccctcc atgctggggt tggatcctcc tgtcttcctc 900 cttgtcccat gaaggccaca ccccccagct ctggggactc ctaggtcact tgggaactac 960 ctgcatcttc agtccccttg aacttctgcc ctctgttcag ggctgacaca agccccacag 1020 gctggggggc tccggttccc tgagggatga gccttcagcc tccctttgta atgctgctcc 1080 tctccactgc ccagcaccat gagttgggtg cagacaccta gaaggagaga cttcttggaa 1140 cgctcatccc ccgctatacc tccccttcct cctgcatctc cccttctttc cttccccctc 1200 aggagagaga aaacttagtg cttccagccc ttcttggagc cttcatggtc caggggtagg 1260 ggccccactg gcctgagcat gccattttga ggggagggta gttgtgccta cttatcccct 1320 ggcagagggg atgccaggac catggacatg aggcttgccc atccctgcca acttacacag 1380 cctgtaccac tgtcccccct tccttggcta ctttgacatg tgcctgctcc tggcatttca 1440 ataaaacccg gcttgggtct gtgtcttgag tgttttttaa aaaaaaaaaa aaa 1493 <210> SEQ ID NO 38 <211> LENGTH: 2930 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 2929276 <400> SEQUENCE: 38 tggagagcgg ggagttgtgt ccaccttgcc gacgtcgcta gccgtggggc tgtcctggga 60 aggcggacgg cgagcgcccg gtgtccgcac tcggccgcct gccgtgcccg tctgcgcccg 120 tgtcatcctc actcgggacg cagggaccgt ttttaaatca caggggcgtg tgtcagcctg 180 ccctaggact tcatgtctat atatttcccc attcactgcc ccgactatct gagatcggcc 240 aagatgactg aggtgatgat gaacacccag cccatggagg agatcggcct cagcccccgc 300 aaggatggcc tttcctacca gatcttccca gacccgtcag attttgaccg ctgctgcaaa 360 ctgaaggacc gtctgccctc catagtggtg gaacccacag aaggggaggt ggagagcggg 420 gagctccggt ggccccctga ggagttcctg gtccaggagg atgagcaaga taactgcgaa 480 gagacagcga aagaaaataa agagcagtag agtccctgtg gactcccatg ggtcatacca 540 gccagcatct gttcctgaac tgtgtttttc ccatcatgac ggaagaagag agtgagccgc 600 aattgttctg aaaatgtcaa acgaggcttc tgttttgcac ctgcagatca ccgagttggt 660 tttcttttct tttcttgcct tttttttttt gaaatttgcc gagcagtgga gccctctgac 720 aatttgcaag gccctctgag aaaggaagct gcttagagcc agggggttag tgggtgaggg 780 gagcgagtgc tgtttttgag atcattatct gaactcaggc agcctagtag aggcagtggt 840 gggattccaa tgggtcttgg tgggtgggag gtggggcatg tgcaaagcaa gcaaggaaca 900 tttggggtaa gaaaacaaac atgaggcaaa agaaaaaata catgttttta agaaaacatt 960 gagcagagaa ctgcagccag gatgcgctca gcagacattc actctggctg ctgggacatc 1020 agaaaacaaa gtcttcatct ctctctccag tttcacccac cccacccttt gctttcattt 1080 caggtgtgtt ggtctatatg acagggagga gagtaaagga gagcaggagc aattggctgc 1140 ctgcaaagcc agctggaggt gaagtgcagg aaaggaaagg tcaccccatt ctactccatg 1200 gcctctctgc tcccagctgt ggtaggctca catagccagt gtgatcggtt tttaagaggc 1260 agtgcttttc agcttttctc cctgatatat ccattttgct tcccagcact ttttaggagt 1320 agtgagagca cttcctgccc ttgttggaag ccccagggtg gacactcagc acgaaggtct 1380 ctcccttaac tgctgccctt ccaagacttg ctcccgagat ggagtgggcg tggtcttcca 1440 ggctggccct tccttctcct caccgccacc ttccctgccc cagccccagc agccatgggt 1500 acatgggtcc ccagctcacc tatggattcc cgccagtctg cccagctgca gtactcacgc 1560 cccatggggg atcttggtct gtttttcttg tgggagccta gtggagagca gacgtggctt 1620 tttatgtgtc ttgttgggga ggtgacttgc atggtgggga caaggctgtc gtggcaacct 1680 tgggatcgag tttgagacta aaggatgtca tgagatccct ggcttctccc catgttgttc 1740 ccggacaagg gcagaaggga ggcatggcaa gggacctctg ctgtccttac tcaacagtgg 1800 tcctcatccc tccccacctc ccactgcttc ctgcaagggc accagttgta tgagaaagtt 1860 ggcctttgga cttaggattt cttattgtag ctaagagcca tctgaagcag caggttgcag 1920 gacaaatgct tcagtccgcc gagagcagta ccgtgtggcc aagaggtgga ctcagagcct 1980 tccttgagct aaactcggcc aaccaaggca cgcagcatgt cccctcaggt ctccagtcag 2040 tccaggttga ccctcagttc tggacgtgtg tatatagctg tatttaatac ctcaaggtca 2100 ttgtggctct ggggatgcca gggcaggagg acgagggtgc gctgtggaca cagcagtccg 2160 cggaattccg ttctgggaag ccaatggtcg ccggcacccc ttgcttcctc cctctgttgt 2220 ctgcctgtgt gacacacatc aatggcaata acttcttcca actcctcgca gaagtgggag 2280 aggccggcag cctgcaccga gaggggcttt cctctctctt gctccccgct tcgttctgtt 2340 ttggctgcag agagtggttc atccatactc tcattccctc gcctcccctt gtggacgggg 2400 gtcttgcctt ttcaattcct gtgttttggt gtcttccctt atctgctacc ctgaatcacc 2460 tgtcctggtc ttgctgtgtg atgggaacat gcttgtaaac tgcgtaacaa atctactttg 2520 tgtatgtgtc tgtttatggg ggtggtttat tatttttgct ggtccctaga ccactttgta 2580 tgaccgtttg cagtctgagc aggccagggg ctgacagcta atgtcaggac cctcagcggt 2640 ggagcctgct ggggggaccc agctgctctt ggacaagtgg ctgagctcct atctggcctc 2700 ctcttttttt tttttttcaa gtaatttgtg tgtatttcta actgattgta ttgaaaaaat 2760 tcctagtatt tcagtaaaaa tgcctgttgt gagatgaacc tcctgtaact tctatctgtt 2820 cttttttgag gctcagggag aaactagcat tttttttttt ccaaactact ttttgtcact 2880 gtgacagttg taaataaagt ttgaaaatgc tttccaaaaa aaaaaaaaaa 2930 <210> SEQ ID NO 39 <211> LENGTH: 4204 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 3033039 <400> SEQUENCE: 39 atgacaatgg atgctctgtt ggctcgattg aaacttctga atccagatga ccttagagaa 60 gaaatcgtca aagccggatt gaaatgtgga cccattacat caactacaag gttcattttt 120 gagaaaaaat tggctcaggc tttactggag caaggaggaa ggctgtcttc tttctaccac 180 catgaggcag gtgtcacagc tctcagccag gacccacaaa ggattttgaa gccagctgaa 240 gggaacccaa ctgatcaggc tggtttttct gaagacagag attttggtta cagtgtgggc 300 ctgaatcctc cagaggagga agctgtgaca tccaagacct gctcggtgcc ccctagtgac 360 accgacacct acagagctgg agcgactgcg tctaaggagc cgcccctgta ctatggggtg 420 tgtccagtgt atgaggacgt cccagcgaga aatgaaagga tctatgttta tgaaaataaa 480 aaggaagcat tgcaagctgt caagatgatc aaagggtccc gatttaaagc tttttctacc 540 agagaagacg ctgagaaatt tgctagagga atttgtgatt atttcccttc tccaagcaaa 600 acgtccttac cactgtctcc tgtgaaaaca gctccactct ttagcaatga caggttgaaa 660 gatggtttgt gcttgtcgga atcagaaaca gtcaacaaag agcgagcgaa cagttacaaa 720 aatccccgca cgcaggacct caccgccaag cttcggaaag ctgtggagaa gggagaggag 780 gacacctttt ctgaccttat ctggagcaac ccccggtatc tgataggctc aggagacaac 840 cccactatcg tgcaggaagg gtgcaggtac aacgtgatgc atgttgctgc caaagagaac 900 caggcttcca tctgccagct gactctggac gtcctggaga accctgactt catgaggctg 960 atgtaccctg atgacgacga ggccatgctg cagaagcgta tccgttacgt ggtggacctg 1020 tacctcaaca cccccgacaa gatgggctat gacacaccgt tgcattttgc ttgtaagttt 1080 ggaaatgcag atgtagtcaa cgtgctttcg tcacaccatt tgattgtaaa aaactcaagg 1140 aataaatatg ataaaacacc tgaagatgta atttgtgaaa gaagcaaaaa taaatctgtg 1200 gaactgaagg agcggatcag agagtattta aagggccact actacgtgcc cctcctgaga 1260 gcggaagaga cttcttctcc agtcatcggg gagctgtggt ccccagacca gacggctgag 1320 gcctctcacg tcagccgcta tggaggcagc cccagagacc cggtactgac cctgagagcc 1380 ttcgcagggc ccctgagtcc agccaaggca gaagattttc gcaagctctg gaaaactcca 1440 cctcgagaga aagcaggctt ccttcaccac gtcaagaagt cggacccgga aagaggcttt 1500 gagagagtgg gaagggagct agctcatgag ctggggtatc cctgggttga atactgggaa 1560 tttctgggct gttttgttga tctgtcttcc caggaaggcc tgcaaagact agaagaatat 1620 ctcacacagc aggaaatagg caaaaaggct caacaagaaa caggagaacg ggaagcctcc 1680 tgccgagata aagccaccac gtctggcagc aattccattt ccgtgagggc gtttctagat 1740 gaagatgaca tgagcttgga agaaataaaa aatcggcaaa atgcagctcg aaataacagc 1800 ccgcccacag tcggtgcttt tggacatacg aggtgcagcg ccttcccctt ggagcaggag 1860 gcagacctca tagaagccgc cgagccggga ggtccacaca gcagcagaaa tgggctctgc 1920 catcctctga atcacagcag gaccctggcg ggcaagagac caaaggcccc ccgtggggag 1980 gaagcccatc tgccacctgt ctcggatttg actgttgagt ttgataaact gaatttgcaa 2040 aatataggac gtagcgtttc caagacacca gatgaaagta caaaaactaa agatcagatc 2100 ctgacttcaa gaatcaatgc agtagaaaga gacttgttag agccttctcc cgcagaccaa 2160 ctcgggaatg gccacaggag gacagaaagt gaaatgtcag ccaggatcgc taaaatgtcc 2220 ttgagtccca gcagccccag gcacgaggat cagctcgagg tcaccaggga accggccagg 2280 cggctcttcc tttttggaga ggagccatca aaactcgatc aggatgtttt ggccgctctt 2340 gaatgtgcag acgtcgaccc ccatcagttc ccggccgtgc acagatggaa gagtgctgtc 2400 ctgtgctact caccctcgga cagacagagt tggcccagtc ccgcggtgaa aggaaggttc 2460 aagtctcagc tgccagatct cagtggccct cacagctaca gtccggggag aaacagcgtg 2520 gctggaagca accccgcaaa gccaggcctg ggcagtcctg ggcgctacag ccccgtgcac 2580 gggagccagc tccgcaggat ggcgcgcctg gctgagcttg ccgccctgta ggcttggcgc 2640 tgggctctcg gtttgttctt catttttaaa gaaggaaggg tcatatgttt attgctaaac 2700 tgtcaaaaag gaatatattc tgattaaatt attactcctc actttgaggg tgtgagaatt 2760 ttagaagatt taaatgttct atataacact tagatttctg atattttgga agaagttaga 2820 agttaatgaa agcaaactca gttaccaatt ttctggaaaa tatccatgtg gtaatggtag 2880 actttttagg tggcaatttc taggtctgaa atatagcaga ggaaagggcg ctgaggcagt 2940 tgcaggcagg cagccctgta cttaccctgt actcacctca tccgacagac gctgtggatg 3000 aggaggggct tggcggaggc gtgagcaccg atgtcccttt gataacctgc actcaccaag 3060 atgaactatt tgccgccctg tcttttcctg ggttgggggg tggcatctga tggtggcaga 3120 gtgcctgttg gttcgcccgt gggtctcatg gttcagacag agggaggtgg acggcaggga 3180 tcagggagcc aggagcgcgc ctcagacttg cagcaaccat tgtgatttgg gttgttcgga 3240 atatttaaat tactgatcag aagatgaaag tagcttttct cttgggaagt cttgcagccc 3300 gtgggagtga taccaggagc aacacagagc tcagcagcgg cgccaaggtg ttccctgttt 3360 cctcagcacg tgagccttca ccgcctgctt cattcaggag ccagtgcagc agtaatacag 3420 tctatacatt gttctgtttt caaatttatc ctgaggcttt gttgagcata aatgattata 3480 cgataaaggt atccgttatt ttggaactca tttcagttgg gatctcctgt atgcagagtg 3540 ttgcatttag aggtttgagt cccatcttgg tttcttgccg tgctgactgt agccttcacc 3600 ttgacttgaa tgaaggtctg tggttggaat gtgtgaggag ccgctgaggt gttcaggagg 3660 tgctgcctgg aggtcggttt cttcctgggt gttacgggca actgctcaca cagttgtttc 3720 tctgtgaaca tttccagtgt ttaatccaaa atgaaaaccc accaatgctt ttgctaactt 3780 cagtgccttt tataaatcat ttttaaattt cctgaacttg ctttttgagg atatacaggg 3840 atattaagta gacgcaggat tgtttttgtt tgtaaaaatt ctgaattgaa actttgtttt 3900 aaaaaaaggc ttctttcttt catatgacaa gagataggtc aggaatattg gaatcaagat 3960 ttaaatgtta aaattcgatt ttgttacaca gggtgtgttc atttgttttg tagcagacaa 4020 gatctagatc ccagacagaa acaacacatg ctattctaaa aagccgcatt ttaaaaggca 4080 ccttggttct caaaagaaat cagaatatgg atattcgtag tgatgatctg ttttctctaa 4140 aatcttacca tattgtctgt atatggttgt aaattcaaat ggaaagtaaa acgttttggc 4200 cctg 4204 <210> SEQ ID NO 40 <211> LENGTH: 2054 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: misc_feature <223> OTHER INFORMATION: Incyte Clone 3039890 <400> SEQUENCE: 40 ggaaggcggt gctctgagaa gccggactac gcggcagcgg ctcttcaaag cggagccggg 60 agtttttgct acagttttcg ccaccatgag tcgcagctat aatgatgagc tgcagttctt 120 ggagaagatc aataaaaact gctggaggat caagaagggc ttcgtgccca acatgcaggt 180 tgaaggtgtt ttctatgtga atgatgctct ggagaaattg atgtttgagg aattaaggaa 240 tgcctgtcga ggtggtggtg ttggtggctt cctgccagcc atgaaacaga ttggcaatgt 300 ggcagccctg cctggaattg ttcatcgatc tattgggctt cctgatgtcc attcaggata 360 tgggtttgct attgggaaca tggcagcctt tgatatgaat gaccctgaag cagtagtatc 420 cccaggtggt gtcgggtttg acatcaactg tggtgtccgc ttgctaagaa ccaatttaga 480 tgaaagtgat gtccagcctg tgaaggagca acttgcccaa gctatgtttg accacattcc 540 tgttggggtg gggtcaaaag gtgtcatccc aatgaatgcc aaagacttgg aggaggcctt 600 ggagatgggg gtggactggt ccttaagaga agggtatgcc tgggctgaag acaaggagca 660 ctgcgaggag tacggaagga tgctgcaggc tgaccccaat aaagtttctg caagggcgaa 720 gaaaagaggc cttcctcagt tggggaccct gggagcaggc aaccattatg cagaaatcca 780 ggttgtggat gagattttca atgagtatgc tgctaaaaaa atgggcatcg accataaggg 840 acaggtgtgt gtgatgatcc acagtggaag cagaggcttg ggccaccaag tagccacaga 900 tgcgctggta gctatggaga aggccatgaa gagagacaag attatagtca atgatcggca 960 gttggcttgt gctcgaatcg cttccccaga gggtcaagac tatctgaagg gaatggcagc 1020 tgctgggaac tatgcctggg tcaaccgctc ttccatgacc ttcttaaccc gtcaggcttt 1080 cgccaaggtc ttcaacacaa cccctgatga cttggaccta catgtgatct atgatgtttc 1140 tcacaacatt gccaaagtgg agcagcatgt ggtggacgga aaggaacgga cactgttagt 1200 acacaggaag ggatccaccc gcgctttccc tcctcaccat cccctcattg ctgttgatta 1260 ccaactcact ggacagccag tgctcattgg tggcaccatg ggaacctgta gttatgttct 1320 tactggcact gaacagggca tgactgagac ctttggaaca acctgtcatg gagcgggccg 1380 tgcattgtcc cgagcaaaat ctcgacgtaa tttagatttc caggatgtct tagacaaatt 1440 ggcagatatg ggaattgcga tccgtgttgc ctcacccaaa ctggttatgg aagaggctcc 1500 tgagtcctat aagaatgtga cagatgtggt aaatacctgc catgatgctg gaatcagcaa 1560 gaaagccatt aaactgagac caattgctgt gatcaaagga tagaaccttg gacagcaggg 1620 ctgcctgaca ccaccaaccc tctctgaagt ggaagtggac tgacatgctc ttctgacatc 1680 agactcaagg cgggacaagt tgcaaagtgt gcagctgtaa ctgctcacgc caaaatggct 1740 gatggggagg ctgctgcttt caggggcccg tgcttgtaaa ataaccttcc aggaagaggc 1800 acattgccca cctttggaaa gggaggaata tgccttctcc ttggttgttc cacagagttt 1860 taggaaaatc tgttagggat gggtagatgt caaactgcct tacgcagtca tactgatctt 1920 tagccatcag attgatcttc ttcacaccaa gctctgttta cattccgaga ggtgtcatga 1980 agaaagttct gttcaataag gttttggaat gtttcaaaaa aaaaaaaaaa ggctggctgc 2040 ttgtgatcta gaac 2054 

What is claimed is:
 1. A purified polypeptide comprising an amino acid sequence selected from the group consisting of: e) an amino acid sequence of SEQ ID NO:1-20, f) a naturally-occurring amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:1-20, c) a biologically active fragment of the polypeptide having the amino acid sequence of SEQ ID NO:1-20, and d) an immunogenic fragment of the polypeptide having the amino acid sequence of SEQ ID NO:1-20.
 2. An isolated polypeptide of claim 1, having an amino acid sequence selected from the group consisting of SEQ ID NO:1-20.
 3. An isolated polynucleotide encoding a polypeptide of claim
 1. 4. An isolated polynucleotide encoding a polypeptide of claim
 2. 5. An isolated polynucleotide of claim 4 comprising a sequence selected from the group consisting of SEQ ID NO:21-40.
 6. A recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide of claim
 3. 7. A cell transformed with a recombinant polynucleotide of claim
 6. 8. A transgenic organism comprising a recombinant polynucleotide of claim
 6. 9. A method for producing a polypeptide of claim 1, the method comprising: a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide, and said recombinant polynucleotide comprises a promoter sequence operably linked to a polynucleotide encoding the polypeptide of claim 1, and b) recovering the polypeptide so expressed.
 10. An isolated polynucleotide selected from the group consisting of: a) a polynucleotide comprising a polynucleotide sequence of SEQ ID NO:21-40, b) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence of SEQ ID NO:21-40, c) a polynucleotide complementary to a polynucleotide of a), d) a polynucleotide complementary to a polynucleotide of b), and e) an RNA equivalent of a)-d).
 11. An isolated polynucleotide comprising at least 60 contiguous nucleotides of a polynucleotide of claim
 10. 12. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 10, the method comprising: a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and, optionally, if present, the amount thereof.
 13. A method of claim 12, wherein the probe comprises at least 60 contiguous nucleotides.
 14. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 10, the method comprising: a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction amplification, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.
 15. A composition comprising a polypeptide of claim 1 and a pharmaceutically acceptable excipient.
 16. A composition of claim 15, wherein the polypeptide has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NO:1-20.
 17. A method for treating a disease or condition associated with decreased expression of FLEXGEM comprising administering to a patient in need of such treatment the composition of claim
 15. 18. A method for screening a compound for effectiveness as an agonist of a polypeptide of claim 1, the method comprising: a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting agonist activity in the sample.
 19. A composition comprising an agonist compound identified by a method of claim 18 and a pharmaceutically acceptable excipient.
 20. A method for treating a disease or condition associated with decreased expression of FLEXGEM, comprising administering to a patient in need of such treatment a composition of claim
 19. 21. A method for screening a compound for effectiveness as an antagonist of a polypeptide of claim 1, the method comprising: a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting antagonist activity in the sample.
 22. A composition comprising an antagonist compound identified by a method of claim 21 and a pharmaceutically acceptable excipient.
 23. A method for treating a disease or condition associated with overexpression of FLEXGEM, comprising administering to a patient in need of such treatment a composition of claim
 22. 24. A method of screening for a compound that specifically binds to the polypeptide of claim 1, said method comprising the steps of: a) combining the polypeptide of claim 1 with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide of claim 1 to the test compound, thereby identifying a compound that specifically binds to the polypeptide of claim
 1. 25. A method of screening for a compound that modulates the activity of the polypeptide of claim 1, said method comprising: a) combining the polypeptide of claim 1 with at least one test compound under conditions permissive for the activity of the polypeptide of claim 1, b) assessing the activity of the polypeptide of claim 1 in the presence of the test compound, and c) comparing the activity of the polypeptide of claim 1 in the presence of the test compound with the activity of the polypeptide of claim 1 in the absence of the test compound, wherein a change in the activity of the polypeptide of claim 1 in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide of claim
 1. 26. A method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a sequence of claim 5, the method comprising: a) exposing a sample comprising the target polynucleotide to a compound, under conditions suitable for the expression of the target polynucleotide, b) detecting altered expression of the target polynucleotide, and c) comparing the expression of the target polynucleotide in the presence of varying amounts of the compound and in the absence of the compound.
 27. A method for assessing toxicity of a test compound, said method comprising: a) treating a biological sample containing nucleic acids with the test compound; b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide of claim 10 under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide comprising a polynucleotide sequence of a polynucleotide of claim 10 or fragment thereof; c) quantifying the amount of hybridization complex; and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.
 28. An isolated antibody which specifically binds to a polypeptide of claim
 1. 29. A diagnostic test for a condition or disease associated with the expression of FLEXGEM in a biological sample comprising the steps of: a) combining the biological sample with an antibody of claim 28, under conditions suitable for the antibody to bind the polypeptide and form an antibody:polypeptide complex; and b) detecting the complex, wherein the presence of the complex correlates with the presence of the polypeptide in the biological sample.
 30. The antibody of claim 28, wherein the antibody is: a) a chimeric antibody, b) a single chain antibody, c) a Fab fragment, d) a F(ab′)₂ fragment, or e) a humanized antibody.
 31. A composition comprising an antibody of claim 28 and an acceptable excipient.
 32. A method of diagnosing a condition or disease associated with the expression of FLEXGEM in a subject, comprising administering to said subject an effective amount of the composition of claim
 31. 33. A composition of claim 31, wherein the antibody is labeled.
 34. A method of diagnosing a condition or disease associated with the expression of FLEXGEM in a subject, comprising administering to said subject an effective amount of the composition of claim
 33. 35. A method of preparing a polyclonal antibody with the specificity of the antibody of claim 28 comprising: a) immunizing an animal with a polypeptide of claim 1, or an immunogenic fragment thereof, under conditions to elicit an antibody response; b) isolating antibodies from said animal; and c) screening the isolated antibodies with the polypeptide, thereby identifying a polyclonal antibody which binds specifically to a polypeptide having an amino acid sequence of a polypeptide of claim
 1. 36. An antibody produced by a method of claim
 35. 37. A composition comprising the antibody of claim 36 and a suitable carrier.
 38. A method of making a monoclonal antibody with the specificity of the antibody of claim 28 comprising: a) immunizing an animal with a polypeptide having an amino acid sequence of a polypeptide of claim 2, or an immunogenic fragment thereof, under conditions to elicit an antibody response; b) isolating antibody producing cells from the animal; c) fusing the antibody producing cells with immortalized cells to form monoclonal antibody-producing hybridoma cells; d) culturing the hybridoma cells; and e) isolating from the culture monoclonal antibody which binds specifically to a polypeptide having an amino acid sequence of a polypeptide of claim
 2. 39. A monoclonal antibody produced by a method of claim
 38. 40. A composition comprising the antibody of claim 39 and a suitable carrier.
 41. The antibody of claim 28, wherein the antibody is produced by screening a Fab expression library.
 42. The antibody of claim 28, wherein the antibody is produced by screening a recombinant immunoglobulin library.
 43. A method for detecting a polypeptide having an amino acid sequence of a polypeptide of claim 2 in a sample, comprising the steps of: a) incubating the antibody of claim 28 with a sample under conditions to allow specific binding of the antibody and the polypeptide; and b) detecting specific binding, wherein specific binding indicates the presence of a polypeptide having an amino acid sequence of a polypeptide of claim 2 in the sample.
 44. A method of purifying a polypeptide having an amino acid sequence of a polypeptide of claim 2 from a sample, the method comprising: a) incubating the antibody of claim 28 with a sample under conditions to allow specific binding of the antibody and the polypeptide; and b) separating the antibody from the sample and obtaining the purified polypeptide having an amino acid sequence of a polypeptide of claim
 2. 45. An isolated polynucleotide comprising a sequence of SEQ ID NO:21.
 46. An isolated polynucleotide comprising a sequence of SEQ ID NO:22.
 47. An isolated polynucleotide comprising a sequence of SEQ ID NO:26.
 48. An isolated polynucleotide comprising a sequence of SEQ ID NO:27.
 49. An isolated polynucleotide comprising a sequence of SEQ ID NO:33. 